Herpes simplex virus vaccine

ABSTRACT

Herpes Simplex Virus (HSV) antigens that elicit an HSV-specific immune response and can be used to treat or prevent HSV infection are provided. Nucleic acid sequences, polypeptides, vectors, and compositions, as well as methods to induce an immune response against HSV, treat or prevent HSV disease, induce a T cell response against HSV, and induce an antibody response against HSV also are provided.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH AND DEVELOPMENT

This invention was made with Government support under Grant Number5R43AI077147-02 awarded by the National Institutes of Health. TheGovernment has certain rights in this invention.

INCORPORATION-BY-REFERENCE OF MATERIAL ELECTRONICALLY SUBMITTED

Incorporated by reference in its entirety herein is a computer-readablenucleotide/amino acid sequence listing submitted concurrently herewithand identified as follows: 1,009,461 bytes ASCII (Text) file named“716196_ST25.txt” created Nov. 24, 2014.

BACKGROUND OF THE INVENTION

Herpes Simplex Virus 2 (HSV-2) is highly infectious and prevalent bothin the United States and worldwide. Population based data have shownthat in the U.S., 17.8% of the general U.S. population has acquiredHSV-2 infection. In some demographic groups such as African Americanwomen, the seroprevalence approaches 60%. Women possess higherseroprevalence rates than men, and HSV-2 seroprevalence rates in manyareas of the world are double that of the U.S. population.

Concomitant with the epidemic of HSV-2 in the U.S. is the increasingprevalence of neonatal HSV-2. An estimated 1,300 cases of neonatal HSVare seen yearly—a higher number of cases than neonatal HIV ever achievedin the U.S. Neonatal HSV, even treated, has a mortality of >15%, and theneurological morbidity among HSV-2 infected infants is an additional30-50% of surviving cases. Case series indicate that 70% of neonatal HSVcases are related to the acquisition of HSV-1 or HSV-2 by the mother inlate pregnancy.

The increasing prevalence of HSV-2 in the adult population has occurreddespite the development and widespread use of antiviral therapy forHSV-2. Antecedent HSV-2 increases the risk of HIV infection by 2-3 fold.Data from Rakai, Uganda, show that on a per contact basis the HSV-2infected person has a 5-7 fold increased rate of HIV-1 acquisition thanthe HSV-2 seronegative person. Mathematical modeling of theepidemiological data has indicated that from ⅓ to ½ of the cases ofHIV-1 in areas of Africa such as Kisumu, Kenya, can be directlyattributed to HSV-2. This effect on HIV is higher for HSV-2 than anyother sexually transmitted illness (STI). HSV-2/HIV co-infected personsappear to be a major “super spreader” of HIV within their communities.In addition, large scale international studies have shown theineffectiveness of antiviral therapy of HSV-2 to reduce HIV-1acquisition and demonstrated the inability of acyclovir to reducetransmission between HIV-1 discordant couples.

The genome of Herpes Simplex Viruses (HSV-1 and HSV-2) contains about 85open reading frames, such that HSV can generate at least 85 uniqueproteins. These genes encode 4 major classes of proteins: (1) thoseassociated with the outermost external lipid bilayer of HSV (theenvelope), (2) the internal protein coat (the capsid), (3) anintermediate complex connecting the envelope with the capsid coat (thetegument), and (4) proteins responsible for replication and infection.

Examples of envelope proteins include UL1 (gL), UL10 (gM), UL20, UL22,UL27 (gB), UL43, UL44 (gC), UL45, UL49A, UL53 (gK), US4 (gG), US5 (gJ),US6 (gD), US7 (gI), US8 (gE), and US10. Examples of capsid proteinsinclude UL6, UL18, UL19, UL35, and UL38. Tegument proteins include UL11,UL13, UL21, UL36, UL37, UL41, UL45, UL46, UL47, UL48, UL49, US9, andUS10. Other HSV proteins include UL2, UL3 UL4, UL5, UL7, UL8, UL9, UL12,UL14, UL15, UL16, UL17, UL23, UL24, UL25, UL26, UL26.5, UL28, UL29,UL30, UL31, UL32, UL33, UL34, UL39, UL40, UL42, UL50, UL51, UL52, UL54,UL55, UL56, US1, US2, US3, US81, US11, US12, ICP0, and ICP4.

Since the envelope (most external portion of an HSV particle) is thefirst to encounter target cells, much of the early HSV-2 vaccinedevelopment work focused on using proteins associated with the envelopeas immunogenic agents. In brief, surface and membraneproteins—glycoprotein D (gD), glycoprotein B (gB), glycoprotein H (gH),glycoprotein L (gL)—as single antigens or in combination with or withoutadjuvants have been tested as possible vaccine antigens. Each was ableto stimulate neutralizing antibody titers and “protect” HSV-2 infectedanimals in challenge models. In humans, all of these vaccines elicitedHSV specific neutralizing antibodies among seronegative and HSV-1seropositive individuals. Neutralizing antibody titers were found to beequal to or 5-10 fold higher than that measured in HSV-2 seropositiveindividuals.

The most promising candidate was glycoprotein D in which multipleclinical trials were conducted, including a very large Phase III trial.Results from this clinical trial showed that, though circulatingneutralizing antibody titers were present and high, only 35% of theseronegative women showed a reduction in HSV-1 acquisition. No benefitwas observed in men. These disappointing findings indicate that thestimulation of neutralizing antibodies is insufficient for an effectiveHSV-2 vaccine. More specifically, these results suggest thatimmunization with envelope proteins for induction of neutralizingantibodies is inadequate to generate an effective HSV vaccine.

There is a need for an effective HSV vaccine for the public healthcontrol of HSV infection.

BRIEF SUMMARY OF THE INVENTION

The invention provides an isolated or purified nucleic acid sequencewith at least 74.5% identity to SEQ ID NO: 1. The invention provides anisolated or purified nucleic acid sequence encoding an amino acidsequence with at least 97% identity to SEQ ID NO: 2. The inventionprovides an isolated or purified nucleic acid sequence with at least82.5% identity to SEQ ID NO: 5.

The invention also provides vectors comprising one or more of theabove-described nucleic acid sequences, such as a vector comprising (i)a nucleic acid sequence with at least 74.5% identity to SEQ ID NO: 1 and(ii) a nucleic acid sequence with at least 82.5% identity to SEQ ID NO:5.

The invention provides polypeptides encoded by the above-describednucleic acid sequences, such as a polypeptide comprising SEQ ID NO: 2.

The invention further provides compositions comprising (i) one or moreof the above-described nucleic acid sequences, vectors, and/orpolypeptides and (ii) a pharmaceutically acceptable carrier, as well asmethods employing the compositions. In particular, the inventionprovides a method of inducing an immune response against HSV in amammal, a method of preventing and/or treating HSV disease in a mammal,a method of inducing a T cell response against HSV in a mammal, and amethod of inducing an antibody response against HSV in a mammalcomprising administering the inventive composition.

The invention also provides for single or multiple administrations(e.g., homologous and/or heterologous administrations, such as primingand boosting compositions) to induce an immune response against HSV in amammal, treat or prevent HSV disease in a mammal, induce an antibodyresponse against HSV in a mammal, and/or induce a T cell responseagainst HSV in a mammal.

BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWING(S)

FIGS. 1A and 1B are graphs showing HSV-specific T cell responsesfollowing a single administration of HSV antigens delivered by anadenoviral vector (1×10⁹ particle units (PU)) to mice as compared toHSV-specific T cell responses following “natural” infection of mice byintravaginal administration (1×10⁶ plaque forming units (PFU)) of HSV-2.The negative control was a single intramuscular administration of FinalFormulation Buffer (FFB) or vehicle. Each of FIGS. 1A and 1B has thepercent of HSV-specific T cells on the y-axis. FIG. 1A demonstrates theT cell response following a single administration of an adenoviralvector comprising SEQ ID NO: 7 (designated LW01) as compared to a“natural” intravaginal infection of a high dose of herpes simplex virus(HSV). FIG. 1B demonstrates the T cell response following a singleadministration of an adenoviral vector comprising SEQ ID NO: 5(designated LW02)) as compared to a “natural” intravaginal infection ofa high dose of HSV. Both graphs show that a single vaccination with HSVantigens (SEQ ID NO: 7 or SEQ ID NO. 5) can produce HSV-specific T celllevels.

FIG. 2 is a graph showing the HSV-specific T cell response followingsingle administration of an adenoviral vector comprising SEQ ID NO: 7(designated LW01), SEQ ID NO: 1 (designated LW11), and SEQ ID NO. 3(designated LW21) as compared to administration of a control (i.e.,vehicle (FFB)). Percent HSV-specific T cells are indicated on they-axis.

FIG. 3 is a graph showing the percent of IFNγ+ CD8+ T cells(HSV-specific T cells) on the y-axis. HSV-specific T cell levels aregreater following two administrations (e.g., homologous or heterologousadministrations) of an adenoviral vector comprising SEQ ID NO: 7 ascompared to a single administration (Ad28 followed by FFB). Miceimmunized with two different adenoviral vectors (e.g., adenoviral vectorserotype 28 (Ad28) followed by a modified adenoviral vector serotype28-designated Ad28H or Ad28 H/F) resulted in higher HSV-specific T celllevels than two administrations of the same adenoviral vector.

FIG. 4 is a graph showing the percent of IFNγ+ CD8+ T cells(HSV-specific T cells) on the y-axis. This graph shows that SEQ ID NO: 5delivered by adenoviral vector serotype 5 (designated Ad5 LW02) andthree other adenoviral vectors (designated GC44 LW02, GC45 LW02, andGC46 LW02) induce high HSV-specific T cell levels when compared tovehicle (FFB) administered mice.

FIGS. 5A and 5B are graphs which illustrate that immunization with anadenoviral vector encoding SEQ ID NO: 5 protects mice against infectionwith HSV. FIG. 5A has mean lesion score on the y-axis and day post HSV-2infection on the x-axis. FIG. 5A shows that a single injection of anadenoviral vector comprising SEQ ID NO: 5 (designated as GC45 LW02)reduces the mean lesion score as compared to the administration ofvehicle (FFB+HSV2) in an HSV mouse infection model. Non HSV-infectedmice were administered phosphate buffer solution (PBS) intravaginallyand served as a negative control for measurement of mean genital lesionscore and as a positive control for survival. FIG. 5B has percentsurvival on the y-axis and days post HSV-2 infection on the x-axis. FIG.5B illustrates that immunization with an adenoviral vector encoding SEQID NO: 5 protects mice infected with HSV.

FIG. 6A is a graph which depicts experimental data illustrating that asingle administration a gorilla adenoviral vector comprising a nucleicacid sequence encoding the inventive UL19 antigen or UL47 antigeninduces robust antigen-specific T cell responses. FIG. 6B is a graphwhich depicts experimental data illustrating that the T cell responsesinduced by a single administration a gorilla adenoviral vectorcomprising a nucleic acid sequence encoding the inventive UL19 antigenor UL47 antigen are more robust than T cell responses induced byinfection with HSV2. Analysis is the mean response±SEM. Statisticalanalysis compared all groups using 2 way ANOVA with Tukey's correctionrepresenting *** p<0.001 and **** p<0.0001.

FIG. 7 is a graph which depicts experimental data illustrating that HSV2infection in ovariectomized female BALB/c mice shows dose dependentsurvival.

FIG. 8 is a graph which depicts experimental data illustrating that asingle administration of a blend of the gorilla adenoviral vectorsGC46.UL19 and GC46.UL47 reduces HSV2 symptoms following HSV2 infectionin mice as compared to administration of either GC46.UL19 or GC46.UL47alone.

FIGS. 9A and 9B are graphs which depict experimental data illustratingthat a single administration of a blend of the gorilla adenoviralvectors GC46.UL19 and GC46.UL47 reduces HSV2 viral shedding in mice asmeasured by qPCR (FIG. 9A) and plaque assay (FIG. 9B).

FIGS. 10A and 10B are graphs which depict experimental data illustratingthat a single administration of a blend of the gorilla adenoviralvectors GC46.UL19 and GC46.UL47 reduces incidence and severity of HSV2lesions in infected guinea pigs.

FIG. 11 is a graph which depicts experimental data illustrating that ahomologous prime/boost immunization method utilizing the GC46.UL19vector or the GC45.UL19 vector results in enhanced T cell responses ascompared to single administration of either vector. In contrast, T cellresponses decreased following homologous boosting with the Ad28HF UL19vector.

FIG. 12A is a graph which depicts experimental data illustrating that ahomologous prime/boost immunization method utilizing the GC46 UL19/UL47adenoviral vector results in enhanced T cell responses as compared tosingle administration of the vector vector, and that the ability toenhance the T cell response is a feature that is applicable to both theinventive UL19 and UL47 antigens.

FIG. 12B is a graph which depicts experimental data illustrating thatsingle administration of the GC46 UL19/UL47 adenoviral vector inducesmucosal T cell responses in mice, and that a second administration ofthe GC46 UL19/UL47 adenoviral vector produces enhanced mucosal T cellresponses as compared to a single administration of the GC46 UL19/UL47vector.

FIG. 13 is a graph which depicts experimental data illustrating that aheterologous prime/boost immunization method utilizing GC45.UL19 as aprime followed by GC46.UL19 as a boost results in enhanced T cellresponses as compared to a single administration of either GC45.UL19 orGC46.UL19 alone.

FIG. 14 is a graph which depicts experimental data illustrating that asingle administration of the GC46.UL19 vector induces elevated durableCD8+ T cell responses for out to 26 weeks post-injection.

DETAILED DESCRIPTION OF THE INVENTION

The invention is predicated, at least in part, on identification of HSVantigens that elicit an HSV (e.g., HSV-1 or HSV-2) specific immuneresponse and/or protect against HSV challenge. The use of the inventiveHSV antigens is therapeutically and prophylactically beneficial topatients with HSV and those at risk of contracting HSV infection.

In particular, a nucleic acid sequence encoding a truncated and modifiedHSV tegument antigen (UL47) that, because of its reduced size, can bemore easily inserted into a variety of molecular delivery systems thanthe larger nucleic acid sequence encoding nontruncated (i.e.,full-length) UL47. The smaller modified sequence (SEQ ID NO: 1)nevertheless produces robust T cell responses comparable to or greaterthan that induced by the nontruncated UL47 antigen. The inventivenucleic acid sequence can be used in a prophylactic and/or therapeuticvaccine for HSV infections.

The inventive (truncated and modified) UL47 antigen is encoded by anucleic acid sequence that is at least 74.5% (e.g., at least 75%, atleast 80%, at least 85%, at least 90%, or at least 95%) identical to SEQID NO: 1. For example, the inventive UL47 nucleic acid sequence can beat least 74.51%, at least 74.57%, at least 74.63%, at least 74.69%, atleast 74.75%, at least 74.81%, at least 74.87%, at least 74.93%, atleast 74.99%, at least 75.04%, at least 75.10%, at least 75.16%, atleast 75.22%, at least 75.28%, at least 75.34%, at least 75.40%, atleast 75.46%, at least 75.52%, at least 75.58%, at least 75.64%, atleast 75.70%, at least 75.76%, at least 75.82%, at least 75.88%, atleast 75.94%, at least 76.00%, at least 76.05%, at least 76.11%, atleast 76.17%, at least 76.23%, at least 76.29%, at least 76.35%, atleast 76.41%, at least 76.47%, at least 76.53%, at least 76.59%, atleast 76.65%, at least 76.71%, at least 76.77%, at least 76.83%, atleast 76.89%, at least 76.95%, at least 77.01%, at least 77.06%, atleast 77.12%, at least 77.18%, at least 77.24%, at least 77.30%, atleast 77.36%, at least 77.42%, at least 77.48%, at least 77.54%, atleast 77.60%, at least 77.66%, at least 77.72%, at least 77.78%, atleast 77.84%, at least 77.90%, at least 77.96%, at least 78.02%, atleast 78.07%, at least 78.13%, at least 78.19%, at least 78.25%, atleast 78.31%, at least 78.37%, at least 78.43%, at least 78.49%, atleast 78.55%, at least 78.61%, at least 78.67%, at least 78.73%, atleast 78.79%, at least 78.85%, at least 78.91%, at least 78.97%, atleast 79.03%, at least 79.08%, at least 79.14%, at least 79.20%, atleast 79.26%, at least 79.32%, at least 79.38%, at least 79.44%, atleast 79.50%, at least 79.56%, at least 79.62%, at least 79.68%, atleast 79.74%, at least 79.80%, at least 79.86%, at least 79.92%, atleast 79.98%, at least 80.04%, at least 80.10%, at least 80.15%, atleast 80.21%, at least 80.27%, at least 80.33%, at least 80.39%, atleast 80.45%, at least 80.51%, at least 80.57%, at least 80.63%, atleast 80.69%, at least 80.75%, at least 80.81%, at least 80.87%, atleast 80.93%, at least 80.99%, at least 81.05%, at least 81.11%, atleast 81.16%, at least 81.22%, at least 81.28%, at least 81.34%, atleast 81.40%, at least 81.46%, at least 81.52%, at least 81.58%, atleast 81.64%, at least 81.70%, at least 81.76%, at least 81.82%, atleast 81.88%, at least 81.94%, at least 82.00%, at least 82.06%, atleast 82.12%, at least 82.17%, at least 82.23%, at least 82.29%, atleast 82.35%, at least 82.41%, at least 82.47%, at least 82.53%, atleast 82.59%, at least 82.65%, at least 82.71%, at least 82.77%, atleast 82.83%, at least 82.89%, at least 82.95%, at least 83.01%, atleast 83.07%, at least 83.13%, at least 83.18%, at least 83.24%, atleast 83.30%, at least 83.36%, at least 83.42%, at least 83.48%, atleast 83.54%, at least 83.60%, at least 83.66%, at least 83.72%, atleast 83.78%, at least 83.84%, at least 83.90%, at least 83.96%, atleast 84.02%, at least 84.08%, at least 84.14%, at least 84.19%, atleast 84.25%, at least 84.31%, at least 84.37%, at least 84.43%, atleast 84.49%, at least 84.55%, at least 84.61%, at least 84.67%, atleast 84.73%, at least 84.79%, at least 84.85%, at least 84.91%, atleast 84.97%, at least 85.03%, at least 85.09%, at least 85.15%, atleast 85.20%, at least 85.26%, at least 85.32%, at least 85.38%, atleast 85.44%, at least 85.50%, at least 85.56%, at least 85.62%, atleast 85.68%, at least 85.74%, at least 85.80%, at least 85.86%, atleast 85.92%, at least 85.98%, at least 86.04%, at least 86.10%, atleast 86.16%, at least 86.22%, at least 86.27%, at least 86.33%, atleast 86.39%, at least 86.45%, at least 86.51%, at least 86.57%, atleast 86.63%, at least 86.69%, at least 86.75%, at least 86.81%, atleast 86.87%, at least 86.93%, at least 86.99%, at least 87.05%, atleast 87.11%, at least 87.17%, at least 87.23%, at least 87.28%, atleast 87.34%, at least 87.40%, at least 87.46%, at least 87.52%, atleast 87.58%, at least 87.64%, at least 87.70%, at least 87.76%, atleast 87.82%, at least 87.88%, at least 87.94%, at least 88.00%, atleast 88.06%, at least 88.12%, at least 88.18%, at least 88.24%, atleast 88.29%, at least 88.35%, at least 88.41%, at least 88.47%, atleast 88.53%, at least 88.59%, at least 88.65%, at least 88.71%, atleast 88.77%, at least 88.83%, at least 88.89%, at least 88.95%, atleast 89.01%, at least 89.07%, at least 89.13%, at least 89.19%, atleast 89.25%, at least 89.30%, at least 89.36%, at least 89.42%, atleast 89.48%, at least 89.54%, at least 89.60%, at least 89.66%, atleast 89.72%, at least 89.78%, at least 89.84%, at least 89.90%, atleast 89.96%, at least 90.02%, at least 90.08%, at least 90.14%, atleast 90.20%, at least 90.26%, at least 90.31%, at least 90.37%, atleast 90.43%, at least 90.49%, at least 90.55%, at least 90.61%, atleast 90.67%, at least 90.73%, at least 90.79%, at least 90.85%, atleast 90.91%, at least 90.97%, at least 91.03%, at least 91.09%, atleast 91.15%, at least 91.21%, at least 91.27%, at least 91.33%, atleast 91.38%, at least 91.44%, at least 91.50%, at least 91.56%, atleast 91.62%, at least 91.68%, at least 91.74%, at least 91.80%, atleast 91.86%, at least 91.92%, at least 91.98%, at least 92.04%, atleast 92.10%, at least 92.16%, at least 92.22%, at least 92.28%, atleast 92.34%, at least 92.39%, at least 92.45%, at least 92.51%, atleast 92.57%, at least 92.63%, at least 92.69%, at least 92.75%, atleast 92.81%, at least 92.87%, at least 92.93%, at least 92.99%, atleast 93.05%, at least 93.11%, at least 93.17%, at least 93.23%, atleast 93.29%, at least 93.35%, at least 93.40%, at least 93.46%, atleast 93.52%, at least 93.58%, at least 93.64%, at least 93.70%, atleast 93.76%, at least 93.82%, at least 93.88%, at least 93.94%, atleast 94.00%, at least 94.06%, at least 94.12%, at least 94.18%, atleast 94.24%, at least 94.30%, at least 94.36%, at least 94.41%, atleast 94.47%, at least 94.53%, at least 94.59%, at least 94.65%, atleast 94.71%, at least 94.77%, at least 94.83%, at least 94.89%, atleast 94.95%, at least 95.01%, at least 95.07%, at least 95.13%, atleast 95.19%, at least 95.25%, at least 95.31%, at least 95.37%, atleast 95.42%, at least 95.48%, at least 95.54%, at least 95.60%, atleast 95.66%, at least 95.72%, at least 95.78%, at least 95.84%, atleast 95.90%, at least 95.96%, at least 96.02%, 96.08%, at least 96.14%,at least 96.20%, at least 96.26%, at least 96.32%, at least 96.38%, atleast 96.43%, at least 96.49%, at least 96.55%, at least 96.61%, atleast 96.67%, at least 96.73%, at least 96.79%, at least 96.85%, atleast 96.91%, at least 96.97%, at least 97.03%, at least 97.09%, atleast 97.15%, at least 97.21%, at least 97.27%, at least 97.33%, atleast 97.39%, at least 97.45%, at least 97.50%, at least 97.56%, atleast 97.62%, at least 97.68%, at least 97.74%, at least 97.80%, atleast 97.86%, at least 97.92%, at least 97.98%, at least 98.04%, atleast 98.10%, at least 98.16%, at least 98.22%, at least 98.28%, atleast 98.34%, at least 98.40%, at least 98.46%, at least 98.51%, atleast 98.57%, at least 98.63%, at least 98.69%, at least 98.75%, atleast 98.81%, at least 98.87%, at least 98.93%, at least 98.99%, atleast 99.05%, at least 99.11%, at least 99.17%, at least 99.23%, atleast 99.29%, at least 99.35%, at least 99.41%, at least 99.47%, atleast 99.52%, at least 99.58%, at least 99.64%, at least 99.70%, atleast 99.76%, at least 99.82%, at least 99.88%, or at least 99.94%identical to SEQ ID NO: 1. In a preferred embodiment, the inventive UL47nucleic acid sequence comprises or consists of SEQ ID NO: 1.

The inventive nucleic acid sequence comprising at least 74.5% identityto SEQ ID NO: 1 preferably is codon-optimized to increase translation ofantigen in infected cells, reduce HSV lesion severity, reduce viralshedding, and protect against HSV infection. The inventive nucleic acidsequence comprising at least 74.5% identity to SEQ ID NO: 1 desirablycontains a C-terminal truncation to enhance antigen processing, andcontains modifications relative to the nucleic acid sequence encodingthe native amino acid sequence (GenBank Accession No. ABX79578), such asto effect amino acid substitutions at positions 291 and 294 in theencoded HSV antigen which preserve reactivity to HSV-1 and HSV-2.Moreover, the nucleic acid sequence comprising at least 74.5% identityto SEQ ID NO: 1 preferably increases the glutamic acid and threonineamino acid content of the translated protein (as indicated by PEST orPEST-like sequences which are signals for rapid protein degradation forMHC recognition).

The invention also provides a polypeptide encoded by the nucleic acidsequence comprising at least 74.5% identity to SEQ ID NO: 1. Inparticular, the invention provides a polypeptide comprising an aminoacid sequence that is at least 97% (e.g., at least 98% or at least 99%)identical to SEQ ID NO: 2. For example, the polypeptide comprises anamino acid sequence that can be at least 97.15%, at least 97.33%, atleast 97.50%, at least 97.68%, at least 97.86%, at least 98.04%, atleast 98.22%, at least 98.40%, at least 98.57%, at least 98.75%, atleast 98.93%, at least 99.11%, at least 99.29%, at least 99.47%, atleast 99.64%, or at least 99.82% identical to SEQ ID NO: 2. In apreferred embodiment, the inventive polypeptide comprises or consists ofthe amino acid sequence of SEQ ID NO: 2.

Nucleic acid or amino acid sequence “identity,” as described herein, canbe determined by comparing a nucleic acid or amino acid sequence ofinterest to a reference nucleic acid or amino acid sequence. The numberof nucleotides or amino acid residues that have been changed and/ormodified (such as, e.g., by point mutations, insertions, or deletions)in the reference sequence so as to result in the sequence of interestare counted. The total number of such changes is subtracted from thetotal length of the sequence of interest, and the difference is dividedby the length of the sequence of interest and expressed as a percentage.A number of mathematical algorithms for obtaining the optimal alignmentand calculating identity between two or more sequences are known andincorporated into a number of available software programs. Examples ofsuch programs include CLUSTAL-W, T-Coffee, and ALIGN (for alignment ofnucleic acid and amino acid sequences), BLAST programs (e.g., BLAST 2.1,BL2SEQ, and later versions thereof) and FASTA programs (e.g., FASTA3x,FASTM, and SSEARCH) (for sequence alignment and sequence similaritysearches). Sequence alignment algorithms also are disclosed in, forexample, Altschul et al., J. Molecular Biol., 215(3): 403-410 (1990);Beigert et al., Proc. Natl. Acad. Sci. USA, 106(10): 3770-3775 (2009);Durbin et al., eds., Biological Sequence Analysis: Probalistic Models ofProteins and Nucleic Acids, Cambridge University Press, Cambridge, UK(2009); Soding, Bioinformatics, 21(7): 951-960 (2005); Altschul et al.,Nucleic Acids Res., 25(17): 3389-3402 (1997); and Gusfield, Algorithmson Strings, Trees and Sequences, Cambridge University Press, CambridgeUK (1997)).

The invention also provides a nucleic acid sequence encoding an HSVcapsid antigen (UL19) that produces robust T cell responses, reduceslesions, protects in an HSV challenge model, and can be used in aprophylactic and/or therapeutic vaccine for HSV infection. Theusefulness of the inventive UL19 as an HSV antigen is unexpected becausethe capsid protein is an internal structural protein and is produced inthe late phases of herpes viral replication and, thus, previously hadnot been considered to be a good antigen for antibody or T cellresponses (see Aubert et al., J. Virol., 75(2), 1013-1030 (2001)). Yet,the UL19 antigen produced by the nucleic acid sequence is veryimmunogenic and produces high T cell levels.

The UL19 antigen is encoded by a nucleic acid sequence that is at least82.5% (e.g., at least 85%, at least 90%, at least 95%) identical to SEQID NO: 5. For example, the inventive UL19 nucleic acid sequence can beat least 82.46%, at least 82.48%, at least 82.51%, at least 82.53%, atleast 82.56%, at least 82.58%, at least 82.61%, at least 82.63%, atleast 82.65%, at least 82.68%, at least 82.70%, at least 82.73%, atleast 82.75%, at least 82.78%, at least 82.80%, at least 82.82%, atleast 82.85%, at least 82.87%, at least 82.90%, at least 82.92%, atleast 82.95%, at least 82.97%, at least 82.99%, at least 83.02%, atleast 83.04%, at least 83.07%, at least 83.09%, at least 83.11%, atleast 83.14%, at least 83.16%, at least 83.19%, at least 83.21%, atleast 83.24%, at least 83.26%, at least 83.28%, at least 83.31%, atleast 83.33%, at least 83.36%, at least 83.38%, at least 83.41%, atleast 83.43%, at least 83.45%, at least 83.48%, at least 83.50%, atleast 83.53%, at least 83.55%, at least 83.58%, at least 83.60%, atleast 83.62%, at least 83.65%, at least 83.67%, at least 83.70%, atleast 83.72%, at least 83.75%, at least 83.77%, at least 83.79%, atleast 83.82%, at least 83.84%, at least 83.87%, at least 83.89%, atleast 83.92%, at least 83.94%, at least 83.96%, at least 83.99%, atleast 84.01%, at least 84.04%, at least 84.06%, at least 84.09%, atleast 84.11%, at least 84.13%, at least 84.16%, at least 84.18%, atleast 84.21%, at least 84.23%, at least 84.26%, at least 84.28%, atleast 84.30%, at least 84.33%, at least 84.35%, at least 84.38%, atleast 84.40%, at least 84.43%, at least 84.45%, at least 84.47%, atleast 84.50%, at least 84.52%, at least 84.55%, at least 84.57%, atleast 84.59%, at least 84.62%, at least 84.64%, at least 84.67%, atleast 84.69%, at least 84.72%, at least 84.74%, at least 84.76%, atleast 84.79%, at least 84.81%, at least 84.84%, at least 84.86%, atleast 84.89%, at least 84.91%, at least 84.93%, at least 84.96%, atleast 84.98%, at least 85.01%, at least 85.03%, at least 85.06%, atleast 85.08%, at least 85.10%, at least 85.13%, at least 85.15%, atleast 85.18%, at least 85.20%, at least 85.23%, at least 85.25%, atleast 85.27%, at least 85.30%, at least 85.32%, at least 85.35%, atleast 85.37%, at least 85.40%, at least 85.42%, at least 85.44%, atleast 85.47%, at least 85.49%, at least 85.52%, at least 85.54%, atleast 85.57%, at least 85.59%, at least 85.61%, at least 85.64%, atleast 85.66%, at least 85.69%, at least 85.71%, at least 85.74%, atleast 85.76%, at least 85.78%, at least 85.81%, at least 85.83%, atleast 85.86%, at least 85.88%, at least 85.90%, at least 85.93%, atleast 85.95%, at least 85.98%, at least 86.00%, at least 86.03%, atleast 86.05%, at least 86.07%, at least 86.10%, at least 86.12%, atleast 86.15%, at least 86.17%, at least 86.20%, at least 86.22%, atleast 86.24%, at least 86.27%, at least 86.29%, at least 86.32%, atleast 86.34%, at least 86.37%, at least 86.39%, at least 86.41%, atleast 86.44%, at least 86.46%, at least 86.49%, at least 86.51%, atleast 86.54%, at least 86.56%, at least 86.58%, at least 86.61%, atleast 86.63%, at least 86.66%, at least 86.68%, at least 86.71%, atleast 86.73%, at least 86.75%, at least 86.78%, at least 86.80%, atleast 86.83%, at least 86.85%, at least 86.88%, at least 86.90%, atleast 86.92%, at least 86.95%, at least 86.97%, at least 87.00%, atleast 87.02%, at least 87.05%, at least 87.07%, at least 87.09%, atleast 87.12%, at least 87.14%, at least 87.17%, at least 87.19%, atleast 87.21%, at least 87.24%, at least 87.26%, at least 87.29%, atleast 87.31%, at least 87.34%, at least 87.36%, at least 87.38%, atleast 87.41%, at least 87.43%, at least 87.46%, at least 87.48%, atleast 87.51%, at least 87.53%, at least 87.55%, at least 87.58%, atleast 87.60%, at least 87.63%, at least 87.65%, at least 87.68%, atleast 87.70%, at least 87.72%, at least 87.75%, at least 87.77%, atleast 87.80%, at least 87.82%, at least 87.85%, at least 87.87%, atleast 87.89%, at least 87.92%, at least 87.94%, at least 87.97%, atleast 87.99%, at least 88.02%, at least 88.04%, at least 88.06%, atleast 88.09%, at least 88.11%, at least 88.14%, at least 88.16%, atleast 88.19%, at least 88.21%, at least 88.23%, at least 88.26%, atleast 88.28%, at least 88.31%, at least 88.33%, at least 88.36%, atleast 88.38%, at least 88.40%, at least 88.43%, at least 88.45%, atleast 88.48%, at least 88.50%, at least 88.52%, at least 88.55%, atleast 88.57%, at least 88.60%, at least 88.62%, at least 88.65%, atleast 88.67%, at least 88.69%, at least 88.72%, at least 88.74%, atleast 88.77%, at least 88.79%, at least 88.82%, at least 88.84%, atleast 88.86%, at least 88.89%, at least 88.91%, at least 88.94%, atleast 88.96%, at least 88.99%, at least 89.01%, at least 89.03%, atleast 89.06%, at least 89.08%, at least 89.11%, at least 89.13%, atleast 89.16%, at least 89.18%, at least 89.20%, at least 89.23%, atleast 89.25%, at least 89.28%, at least 89.30%, at least 89.33%, atleast 89.35%, at least 89.37%, at least 89.40%, at least 89.42%, atleast 89.45%, at least 89.47%, at least 89.50%, at least 89.52%, atleast 89.54%, at least 89.57%, at least 89.59%, at least 89.62%, atleast 89.64%, at least 89.67%, at least 89.69%, at least 89.71%, atleast 89.74%, at least 89.76%, at least 89.79%, at least 89.81%, atleast 89.84%, at least 89.86%, at least 89.88%, at least 89.91%, atleast 89.93%, at least 89.96%, at least 89.98%, at least 90.00%, atleast 90.03%, at least 90.05%, at least 90.08%, at least 90.10%, atleast 90.13%, at least 90.15%, at least 90.17%, at least 90.20%, atleast 90.22%, at least 90.25%, at least 90.27%, at least 90.30%, atleast 90.32%, at least 90.34%, at least 90.37%, at least 90.39%, atleast 90.42%, at least 90.44%, at least 90.47%, at least 90.49%, atleast 90.51%, at least 90.54%, at least 90.56%, at least 90.59%, atleast 90.61%, at least 90.64%, at least 90.66%, at least 90.68%, atleast 90.71%, at least 90.73%, at least 90.76%, at least 90.78%, atleast 90.81%, at least 90.83%, at least 90.85%, at least 90.88%, atleast 90.90%, at least 90.93%, at least 90.95%, at least 90.98%, atleast 91.00%, at least 91.02%, at least 91.05%, at least 91.07%, atleast 91.10%, at least 91.12%, at least 91.15%, at least 91.17%, atleast 91.19%, at least 91.22%, at least 91.24%, at least 91.27%, atleast 91.29%, at least 91.31%, at least 91.34%, at least 91.36%, atleast 91.39%, at least 91.41%, at least 91.44%, at least 91.46%, atleast 91.48%, at least 91.51%, at least 91.53%, at least 91.56%, atleast 91.58%, at least 91.61%, at least 91.63%, at least 91.65%, atleast 91.68%, at least 91.70%, at least 91.73%, at least 91.75%, atleast 91.78%, at least 91.80%, at least 91.82%, at least 91.85%, atleast 91.87%, at least 91.90%, at least 91.92%, at least 91.95%, atleast 91.97%, at least 91.99%, at least 92.02%, at least 92.04%, atleast 92.07%, at least 92.09%, at least 92.12%, at least 92.14%, atleast 92.16%, at least 92.19%, at least 92.21%, at least 92.24%, atleast 92.26%, at least 92.29%, at least 92.31%, at least 92.33%, atleast 92.36%, at least 92.38%, at least 92.41%, at least 92.43%, atleast 92.46%, at least 92.48%, at least 92.50%, at least 92.53%, atleast 92.55%, at least 92.58%, at least 92.60%, at least 92.62%, atleast 92.65%, at least 92.67%, at least 92.70%, at least 92.72%, atleast 92.75%, at least 92.77%, at least 92.79%, at least 92.82%, atleast 92.84%, at least 92.87%, at least 92.89%, at least 92.92%, atleast 92.94%, at least 92.96%, at least 92.99%, at least 93.01%, atleast 93.04%, at least 93.06%, at least 93.09%, at least 93.11%, atleast 93.13%, at least 93.16%, at least 93.18%, at least 93.21%, atleast 93.23%, at least 93.26%, at least 93.28%, at least 93.30%, atleast 93.33%, at least 93.35%, at least 93.38%, at least 93.40%, atleast 93.43%, at least 93.45%, at least 93.47%, at least 93.50%, atleast 93.52%, at least 93.55%, at least 93.57%, at least 93.60%, atleast 93.62%, at least 93.64%, at least 93.67%, at least 93.69%, atleast 93.72%, at least 93.74%, at least 93.77%, at least 93.79%, atleast 93.81%, at least 93.84%, at least 93.86%, at least 93.89%, atleast 93.91%, at least 93.93%, at least 93.96%, at least 93.98%, atleast 94.01%, at least 94.03%, at least 94.06%, at least 94.08%, atleast 94.10%, at least 94.13%, at least 94.15%, at least 94.18%, atleast 94.20%, at least 94.23%, at least 94.25%, at least 94.27%, atleast 94.30%, at least 94.32%, at least 94.35%, at least 94.37%, atleast 94.40%, at least 94.42%, at least 94.44%, at least 94.47%, atleast 94.49%, at least 94.52%, at least 94.54%, at least 94.57%, atleast 94.59%, at least 94.61%, at least 94.64%, at least 94.66%, atleast 94.69%, at least 94.71%, at least 94.74%, at least 94.76%, atleast 94.78%, at least 94.81%, at least 94.83%, at least 94.86%, atleast 94.88%, at least 94.91%, at least 94.93%, at least 94.95%, atleast 94.98%, at least 95.00%, at least 95.03%, at least 95.05%, atleast 95.08%, at least 95.10%, at least 95.12%, at least 95.15%, atleast 95.17%, at least 95.20%, at least 95.22%, at least 95.25%, atleast 95.27%, at least 95.29%, at least 95.32%, at least 95.34%, atleast 95.37%, at least 95.39%, at least 95.41%, at least 95.44%, atleast 95.46%, at least 95.49%, at least 95.51%, at least 95.54%, atleast 95.56%, at least 95.58%, at least 95.61%, at least 95.63%, atleast 95.66%, at least 95.68%, at least 95.71%, at least 95.73%, atleast 95.75%, at least 95.78%, at least 95.80%, at least 95.83%, atleast 95.85%, at least 95.88%, at least 95.90%, at least 95.92%, atleast 95.95%, at least 95.97%, at least 96.00%, at least 96.02%, atleast 96.05%, at least 96.07%, at least 96.09%, at least 96.12%, atleast 96.14%, at least 96.17%, at least 96.19%, at least 96.22%, atleast 96.24%, at least 96.26%, at least 96.29%, at least 96.31%, atleast 96.34%, at least 96.36%, at least 96.39%, at least 96.41%, atleast 96.43%, at least 96.46%, at least 96.48%, at least 96.51%, atleast 96.53%, at least 96.56%, at least 96.58%, at least 96.60%, atleast 96.63%, at least 96.65%, at least 96.68%, at least 96.70%, atleast 96.72%, at least 96.75%, at least 96.77%, at least 96.80%, atleast 96.82%, at least 96.85%, at least 96.87%, at least 96.89%, atleast 96.92%, at least 96.94%, at least 96.97%, at least 96.99%, atleast 97.02%, at least 97.04%, at least 97.06%, at least 97.09%, atleast 97.11%, at least 97.14%, at least 97.16%, at least 97.19%, atleast 97.21%, at least 97.23%, at least 97.26%, at least 97.28%, atleast 97.31%, at least 97.33%, at least 97.36%, at least 97.38%, atleast 97.40%, at least 97.43%, at least 97.45%, at least 97.48%, atleast 97.50%, at least 97.53%, at least 97.55%, at least 97.57%, atleast 97.60%, at least 97.62%, at least 97.65%, at least 97.67%, atleast 97.70%, at least 97.72%, at least 97.74%, at least 97.77%, atleast 97.79%, at least 97.82%, at least 97.84%, at least 97.87%, atleast 97.89%, at least 97.91%, at least 97.94%, at least 97.96%, atleast 97.99%, at least 98.01%, at least 98.03%, at least 98.06%, atleast 98.08%, at least 98.11%, at least 98.13%, at least 98.16%, atleast 98.18%, at least 98.20%, at least 98.23%, at least 98.25%, atleast 98.28%, at least 98.30%, at least 98.33%, at least 98.35%, atleast 98.37%, at least 98.40%, at least 98.42%, at least 98.45%, atleast 98.47%, at least 98.50%, at least 98.52%, at least 98.54%, atleast 98.57%, at least 98.59%, at least 98.62%, at least 98.64%, atleast 98.67%, at least 98.69%, at least 98.71%, at least 98.74%, atleast 98.76%, at least 98.79%, at least 98.81%, at least 98.84%, atleast 98.86%, at least 98.88%, at least 98.91%, at least 98.93%, atleast 98.96%, at least 98.98%, at least 99.01%, at least 99.03%, atleast 99.05%, at least 99.08%, at least 99.10%, at least 99.13%, atleast 99.15%, at least 99.18%, at least 99.20%, at least 99.22%, atleast 99.25%, at least 99.27%, at least 99.30%, at least 99.32%, atleast 99.34%, at least 99.37%, at least 99.39%, at least 99.42%, atleast 99.44%, at least 99.47%, at least 99.49%, at least 99.51%, atleast 99.54%, at least 99.56%, at least 99.59%, at least 99.61%, atleast 99.64%, at least 99.66%, at least 99.68%, at least 99.71%, atleast 99.73%, at least 99.76%, at least 99.78%, at least 99.81%, atleast 99.83%, at least 99.85%, at least 99.88%, at least 99.90%, atleast 99.93%, at least 99.95%, or at least 99.98% identical to SEQ IDNO: 5. In a preferred embodiment, the inventive nucleic acid sequencecomprises or consists of SEQ ID NO: 5.

The inventive nucleic acid sequence comprising at least 82.5% identityto SEQ ID NO: 5 preferably is codon-optimized to increase translation ofantigen in infected cells, reduce HSV lesion severity, reduce viralshedding, and protect against HSV infection. The nucleic acid sequencecomprising at least 82.5% identity to SEQ ID NO: 5 preferably encodes apolypeptide comprising SEQ ID NO: 6 (corresponding to GenBank AccessionNo. CAB06743.1).

The inventive nucleic acids described herein can comprise DNA or RNA,and can be single or double stranded. Furthermore, the nucleic acids cancomprise nucleotide analogues or derivatives (e.g., inosine orphosphorothioate nucleotides and the like). Each nucleic acid can encodea polypeptide alone or as part of a fusion protein. For example, thenucleic acids of the invention can encode a fusion protein comprisingHSV antigen(s) and ubiquitin.

In one embodiment, the inventive nucleic acid sequence comprising atleast 74.5% identity to SEQ ID NO: 1 also comprises a nucleic acidsequence that encodes ubiquitin. For example, the inventive nucleic acidsequence can comprise SEQ ID NO: 3, and the resulting polypeptide cancomprise SEQ ID NO: 4.

Each nucleic acid encoding a polypeptide can be provided as part of aconstruct comprising the nucleic acid and elements that enable deliveryof the nucleic acid to a cell, and/or expression of the nucleic acid ina cell. Such elements include, for example, expression vectors,promoters, and transcription and/or translation sequences. Suitablevectors, promoters, transcription/translation sequences, and otherelements, as well as methods of preparing such nucleic acids andconstructs, are known in the art (e.g., Sambrook et al., MolecularCloning: A Laboratory Manual, 3^(rd) ed., Cold Spring Harbor Press, ColdSpring Harbor, N.Y. (2001); and Ausubel et al., Current Protocols inMolecular Biology, Greene Publishing Associates and John Wiley & Sons,New York (1994)).

The inventive polypeptide can be prepared by any method, such as bysynthesizing the polypeptide or by expressing a nucleic acid encoding anappropriate amino acid sequence in a cell and harvesting the peptidefrom the cell or media containing the cell. A combination of suchmethods also can be used. Methods of de novo synthesizing polypeptidesand methods of recombinantly producing polypeptides are known in the art(see, e.g., Chan et al., Fmoc Solid Phase Peptide Synthesis, OxfordUniversity Press, Oxford, United Kingdom (2005); Reid, R. (ed.), Peptideand Protein Drug Analysis, Marcel Dekker, Inc. (2000); Westwood et al.(ed.), Epitope Mapping, Oxford University Press, Oxford, United Kingdom(2000); Sambrook et al., Molecular Cloning: A Laboratory Manual, 3^(rd)ed., Cold Spring Harbor Press, Cold Spring Harbor, N.Y. (2001); andAusubel et al., Current Protocols in Molecular Biology, GreenePublishing Associates and John Wiley & Sons, New York (1994)).

The invention provides a vector comprising one or more nucleic acidsequences comprising or consisting of a wild-type or inventive UL19and/or wild-type or inventive UL47 nucleic acid sequence(s).Additionally, the invention also provides a composition comprisingmultiple vectors, each of which comprises or consists of one or morenucleic acid sequences comprising the wild-type or inventive UL19 and/orwild-type or inventive UL47 nucleic acid sequence(s). Theabove-described vectors also can comprise one or more nucleic acidsequences encoding one or more additional antigens (e.g., HSV antigens),though the vectors comprising the wild-type UL19 and/or wild-type UL47nucleic acid sequences desirably do not include any other HSV antigennucleic acid sequences.

An “antigen” is a molecule that triggers an immune response in a mammal.An “immune response” can entail, for example, antibody production and/orthe activation of immune effector cells. An antigen in the context ofthe invention can comprise any subunit, fragment, or epitope of anyproteinaceous or non-proteinaceous (e.g., carbohydrate or lipid)molecule that provokes an immune response in a mammal. By “epitope” ismeant a sequence of an antigen that is recognized by an antibody or anantigen receptor. Epitopes also are referred to in the art as “antigenicdeterminants.” The antigen can be a protein or peptide of viral,bacterial, parasitic, fungal, protozoan, prion, cellular, orextracellular origin, which provokes an immune response in a mammal,preferably leading to protective immunity.

Any vector can be employed in the context of the invention, includingviral and non-viral vectors. Examples of suitable viral vectors include,but are not limited to, retroviral vectors, adeno-associated virusvectors, poxviral vectors (e.g., vaccinia virus vectors), herpesvirusvectors, parainfluenza-RSV chimeric vectors (PIV-RSV), adenoviralvectors, poliovirus, alphavirus, baculovirus, and Sindbis virus.Examples of suitable non-viral vectors include, but are not limited to,plasmids (e.g., DNA plasmids), yeast (e.g., Saccharomyces), liposomes,nanoparticles, and molecular conjugates (e.g., transferrin). When thevector is a plasmid (e.g., DNA plasmid), the plasmid can be complexedwith adjuvants, such as CpG or polymeric adjuvants.

Preferably, the vector is an adenoviral vector. Adenovirus from variousorigins, subtypes, or mixture of subtypes can be used as the source ofthe viral genome for the adenoviral vector.

Non-human adenovirus (e.g., ape, simian, avian, canine, ovine, or bovineadenoviruses) can be used to generate the adenoviral vector (i.e., as asource of the adenoviral genome for the adenoviral vector). In oneembodiment, a non-human primate adenovirus (e.g., ape, simian, ofgorilla) can be used to generate the adenoviral vector. For example, theadenoviral vector can be based on a simian adenovirus, including bothnew world and old world monkeys (see, e.g., Virus Taxonomy: VIIIthReport of the International Committee on Taxonomy of Viruses (2005)).The phylogeny of adenoviruses that infect primates is disclosed in,e.g., Roy et al., PLoS Pathog., 5(7): e100050.doi:10.1371/journal.ppat.1000503 (2009). For instance, a simianadenovirus can be of serotype 1, 3, 6, 7, 11, 16, 18, 19, 20, 27, 33,38, 39, 48, 49, or 50, or any other simian adenoviral serotype. Othernon-human adenoviruses which can be used in the invention includenon-human primate adenoviruses that are genetically and/orphenotypically similar to or distinct from group C human adenoviruses.

A gorilla adenovirus can be used as the source of the adenoviral genomefor the adenoviral vector. There are four widely recognized gorillasubspecies within the two species of Eastern Gorilla (Gorilla beringei)and Western Gorilla (Gorilla gorilla). The Western Gorilla speciesincludes the subspecies Western Lowland Gorilla (Gorilla gorillagorilla) and Cross River Gorilla (Gorilla gorilla diehli). The EasternGorilla species includes the subspecies Mountain Gorilla (Gorillaberingei beringei) and Eastern Lowland Gorilla (Gorilla beringeigraueri) (see, e.g., Wilson and Reeder, eds., Mammalian Species of theWorld, 3rd ed., Johns Hopkins University Press, Baltimore, Md. (2005)).The adenoviral vector can be based on an adenovirus isolated from any ofthe aforementioned subspecies. Preferably, the adenoviral vector isbased on an adenovirus isolated from Mountain Gorilla (Gorilla beringeiberingei) or Eastern Lowland Gorilla (Gorilla beringei graueri).

Gorilla adenovirus used as the source of the adenoviral genome can havea nucleic acid sequence that is at least 70% (e.g., at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, at least 96%, atleast 97%, at least 98%, at least 99%, or 100%) identical to the nucleicacid sequence of, for example, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO:11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ IDNO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25,SEQ ID NO: 26, SEQ ID NO: 27, or SEQ ID NO: 28. In particular, theinventive nucleic acid sequence(s) encoding one or more HSV antigensand/or other antigens and/or adjuvants can be inserted into anadenoviral vector described in U.S. Patent Application Nos. 61/543,638,61/543,652, and 61/543,661.

A human adenovirus can be used as the source of the viral genome for theadenoviral vector. For instance, an adenovirus can be of subgroup A(e.g., serotypes 12, 18, and 31), subgroup B (e.g., serotypes 3, 7, 11,14, 16, 21, 34, 35, and 50), subgroup C (e.g., serotypes 1, 2, 5, and6), subgroup D (e.g., serotypes 8, 9, 10, 13, 15, 17, 19, 20, 22-30, 32,33, 36-39, and 42-48), subgroup E (e.g., serotype 4), subgroup F (e.g.,serotypes 40 and 41), an unclassified serogroup (e.g., serotypes 49 and51), or any other adenoviral serogroup or serotype. Adenoviral serotypes1 through 51 (i.e., Ad1 through Ad51) are available from the AmericanType Culture Collection (ATCC, Manassas, Va.). Non-group C adenoviralvectors, methods of producing non-group C adenoviral vectors, andmethods of using non-group C adenoviral vectors are disclosed in, forexample, U.S. Pat. Nos. 5,801,030, 5,837,511, and 5,849,561, andInternational Patent Application Publications WO 1997/012986 and WO1998/053087.

The adenoviral vector can comprise a composition of subtypes and therebybe a “chimeric” adenoviral vector. A chimeric adenoviral vector cancomprise an adenoviral genome that is derived from two or more (e.g., 2,3, 4, etc.) different adenovirus serotypes. In the context of theinvention, a chimeric adenoviral vector can comprise approximatelydifferent or equal amounts of the genome of each of the two or moredifferent adenovirus serotypes. For example, when the chimericadenoviral vector genome is comprised of the genomes of two differentadenovirus serotypes, the chimeric adenoviral vector genome preferablycomprises no more than about 99% (e.g., no more than about 95%, no morethan about 85%, no more than about 80%, no more than about 75%, no morethan about 60%, no more than about 65%, or no more than about 50%) ofthe genome of one of the adenovirus serotypes, with the remainder of thechimeric adenovirus genome being derived from the genome of the otheradenovirus serotype.

In one embodiment, the invention provides a serotype 28 adenovector(Ad28) that contains the hexon and/or fiber (e.g., knob) from adifferent serotype (e.g., a low seroprevalence human adenoviral vector,such as human serotype 45 adenovirus). A description of the viral genomeof such a modified Ad28 vector is set forth in Example 1.

The adenoviral vector for use in the invention can bereplication-competent, conditionally-replicating, orreplication-deficient. Preferably, the adenovirus or adenoviral vectoris replication-deficient, such that the replication-deficient adenovirusor adenoviral vector requires complementation of at least onereplication-essential gene function of one or more regions of theadenoviral genome for propagation (e.g., to form adenoviral vectorparticles).

The replication-deficient adenovirus or adenoviral vector can bemodified in any suitable manner to cause the deficiencies in the one ormore replication-essential gene functions in one or more regions of theadenoviral genome for propagation. The complementation of thedeficiencies in the one or more replication-essential gene functions ofone or more regions of the adenoviral genome refers to the use ofexogenous means to provide the deficient replication-essential genefunctions. Such complementation can be effected in any suitable manner,for example, by using complementing cells and/or exogenous DNA (e.g.,helper adenovirus) encoding the disrupted replication-essential genefunctions.

A deficiency in a gene function or genomic region, as used herein, isdefined as a disruption (e.g., deletion) of sufficient genetic materialof the adenoviral genome to obliterate or impair the function of thegene (e.g., such that the function of the gene product is reduced by atleast about 2-fold, 5-fold, 10-fold, 20-fold, 30-fold, or 50-fold) whosenucleic acid sequence was disrupted (e.g., deleted) in whole or in part.Deletion of an entire gene region often is not required for disruptionof a replication-essential gene function. However, for the purpose ofproviding sufficient space in the adenoviral genome for one or moretransgenes, removal of a majority of one or more gene regions may bedesirable. While deletion of genetic material is preferred, mutation ofgenetic material by addition or substitution also is appropriate fordisrupting gene function. Replication-essential gene functions are thosegene functions that are required for adenovirus replication (e.g.,propagation) and are encoded by, for example, the adenoviral earlyregions (e.g., the E1, E2, and E4 regions), late regions (e.g., the L1,L2, L3, L4, and L5 regions), genes involved in viral packaging (e.g.,the IVa2 gene), and virus-associated RNAs (e.g., VA-RNA-1 and/orVA-RNA-2).

The adenovirus or adenoviral vector can be deficient in one or morereplication-essential gene functions of only the early regions (i.e.,E1-E4 regions) of the adenoviral genome, only the late regions (i.e.,L1-L5 regions) of the adenoviral genome, both the early and late regionsof the adenoviral genome, or all adenoviral genes (i.e., a high capacityadenovector (HC-Ad). See Morsy et al., Proc. Natl. Acad. Sci. USA, 95:965-976 (1998); Chen et al., Proc. Natl. Acad. Sci. USA, 94: 1645-1650(1997); and Kochanek et al., Hum. Gene Ther., 10: 2451-2459 (1999).Examples of replication-deficient adenoviral vectors are disclosed inU.S. Pat. Nos. 5,837,511; 5,851,806; 5,994,106; 6,127,175; 6,482,616;and 7,195,896, and International Patent Application Publications WO1994/028152, WO 1995/002697, WO 1995/016772, WO 1995/034671, WO1996/022378, WO 1997/012986, WO 1997/021826, and WO 2003/022311.

The early regions of the adenoviral genome include the E1, E2, E3, andE4 regions. The late regions of the adenoviral genome include the L1,L2, L3, L4, and L5 regions. The adenovirus or adenoviral vector also canhave a mutation in the major late promoter (MLP), as discussed inInternational Patent Application Publication WO 2000/000628, which canrender the adenovirus or adenoviral vector replication-deficient ifdesired.

The E1 region comprises the E1A and E1B subregions, and one or moredeficiencies in replication-essential gene functions in the E1 regioncan include one or more deficiencies in replication-essential genefunctions in either or both of the E1A and E1B subregions, therebyrequiring complementation of the deficiency in the E1A subregion and/orthe E1B subregion of the adenoviral genome for the adenovirus oradenoviral vector to propagate (e.g., to form adenoviral vectorparticles).

The E2 region comprises the E2A and E2B subregions, and one or moredeficiencies in replication-essential gene functions in the E2 regioncan include one or more deficiencies in replication-essential genefunctions in either or both of the E2A and E2B subregions, therebyrequiring complementation of the deficiency in the E2A subregion and/orthe E2B subregion of the adenoviral genome for the adenovirus oradenoviral vector to propagate (e.g., to form adenoviral vectorparticles).

The E3 region does not include any replication-essential gene functions,such that a deletion of the E3 region in part or in whole does notrequire complementation of any gene functions in the E3 region for theadenovirus or adenoviral vector to propagate (e.g., to form adenoviralvector particles). In the context of the invention, the E3 region isdefined as the region that initiates with the open reading frame thatencodes a protein with high homology to the 12.5K protein from the E3region of human adenovirus 5 (NCBI reference sequence AP_000218) andends with the open reading frame that encodes a protein with highhomology to the 14.7K protein from the E3 region of human adenovirus 5(NCBI reference sequence AP_000224.1). The E3 region may be deleted inwhole or in part, or retained in whole or in part. The size of thedeletion may be tailored so as to retain an adenovirus or adenoviralvector whose genome closely matches the optimum genome packaging size. Alarger deletion will accommodate the insertion of larger heterologousnucleic acid sequences in the adenovirus or adenoviral genome. In oneembodiment of the invention, the L4 polyadenylation signal sequences,which reside in the E3 region, are retained.

The E4 region comprises multiple open reading frames (ORFs). Anadenovirus or adenoviral vector with a deletion of all of the openreading frames of the E4 region except ORF6, and in some cases ORF3,does not require complementation of any gene functions in the E4 regionfor the adenovirus or adenoviral vector to propagate (e.g., to formadenoviral vector particles). Conversely, an adenovirus or adenoviralvector with a disruption or deletion of ORF6, and in some cases ORF3, ofthe E4 region (e.g., with a deficiency in a replication-essential genefunction based in ORF6 and/or ORF3 of the E4 region), with or without adisruption or deletion of any of the other open reading frames of the E4region or the native E4 promoter, polyadenylation sequence, and/or theright-side inverted terminal repeat (ITR), requires complementation ofthe deficiency in the E4 region (specifically, of ORF6 and/or ORF3 ofthe E4 region) for the adenovirus or adenoviral vector to propagate(e.g., to form adenoviral vector particles).

The one or more regions of the adenoviral genome that contain one ormore deficiencies in replication-essential gene functions desirably areone or more early regions of the adenoviral genome, i.e., the E1, E2,and/or E4 regions, optionally with the deletion in part or in whole ofthe E3 region. In other words, the adenoviral vector requires, at most,complementation of a deficiency in one or more early regions of theadenoviral genome for propagation.

The replication-deficient adenovirus or adenoviral vector also can haveone or more mutations as compared to the wild-type adenovirus (e.g., oneor more deletions, insertions, and/or substitutions) in the adenoviralgenome that do not inhibit viral replication in host cells. Thus, inaddition to one or more deficiencies in replication-essential genefunctions, the adenovirus or adenoviral vector can be deficient in otherrespects that are not replication-essential. For example, the adenovirusor adenoviral vector can have a partial or entire deletion of theadenoviral early region known as the E3 region, which is not essentialfor propagation of the adenovirus or adenoviral genome.

In one embodiment, the adenovirus or adenoviral vector isreplication-deficient and requires, at most, complementation of the E1region or the E4 region of the adenoviral genome, for propagation (e.g.,to form adenoviral vector particles). Thus, the replication-deficientadenovirus or adenoviral vector requires complementation of at least onereplication-essential gene function of the E1A subregion and/or the E1Bregion of the adenoviral genome (denoted an E1-deficient adenoviralvector) or the E4 region of the adenoviral genome (denoted anE4-deficient adenoviral vector) for propagation (e.g., to formadenoviral vector particles). The adenovirus or adenoviral vector can bedeficient in at least one replication-essential gene function (desirablyall replication-essential gene functions) of the E1 region of theadenoviral genome and at least one gene function of the nonessential E3region of the adenoviral genome (denoted an E1/E3-deficient adenoviralvector). Such an adenoviral vector requires, at most, complementation ofa deficiency in the E1 region of the adenoviral genome for propagation.The adenovirus or adenoviral vector can be deficient in at least onereplication-essential gene function (desirably all replication-essentialgene functions) of the E4 region of the adenoviral genome and at leastone gene function of the nonessential E3 region of the adenoviral genome(denoted an E3/E4-deficient adenoviral vector). Such an adenoviralvector requires, at most, complementation of a deficiency in the E4region of the adenoviral genome for propagation.

In one embodiment, the adenovirus or adenoviral vector isreplication-deficient and requires, at most, complementation of the E2region, preferably the E2A subregion, of the adenoviral genome, forpropagation (e.g., to form adenoviral vector particles). Thus, thereplication-deficient adenovirus or adenoviral vector requirescomplementation of at least one replication-essential gene function ofthe E2A subregion of the adenoviral genome (denoted an E2A-deficientadenoviral vector) for propagation (e.g., to form adenoviral vectorparticles). The adenovirus or adenoviral vector can be deficient in atleast one replication-essential gene function (desirably allreplication-essential gene functions) of the E2A region of theadenoviral genome and at least one gene function of the nonessential E3region of the adenoviral genome (denoted an E2A/E3-deficient adenoviralvector). Such an adenoviral vector requires, at most, complementation ofa deficiency in the E2A region of the adenoviral genome for propagation.

In one embodiment, the adenovirus or adenoviral vector requirescomplementation of the E1 and E2 (e.g., E2A) regions of the adenoviralgenome for complementation (denoted an E1/E2-deficient adenoviralvector), wherein the adenovirus or adenoviral vector also can bedeficient in at least one gene function of the E3 region (denoted anE1/E2/E3-deficient adenoviral vector). Such an adenoviral vectorrequires, at most, complementation of a deficiency in the E1 region anda deficiency in the E2 region of the adenoviral genome for propagation.

In one embodiment, the adenovirus or adenoviral vector isreplication-deficient and requires, at most, complementation of the E1and E4 regions of the adenoviral genome for propagation (e.g., to formadenoviral vector particles). Thus, the replication-deficient adenovirusor adenoviral vector requires complementation of at least onereplication-essential gene function of both the E1 and E4 regions of theadenoviral genome (denoted an E1/E4-deficient adenoviral vector) forpropagation (e.g., to form adenoviral vector particles). The adenovirusor adenoviral vector can be deficient in at least onereplication-essential gene function (desirably all replication-essentialgene functions) of the E1 region of the adenoviral genome, at least onereplication-essential gene function of the E4 region of the adenoviralgenome, and at least one gene function of the nonessential E3 region ofthe adenoviral genome (denoted an E1/E3/E4-deficient adenoviral vector).

In a preferred embodiment, the adenovirus or adenoviral vector requires,at most, complementation of a deficiency in the E1 region of theadenoviral genome for propagation, and does not require complementationof any other deficiency of the adenoviral genome for propagation. Inanother preferred embodiment, the adenovirus or adenoviral vectorrequires, at most, complementation of a deficiency in both the E1 and E4regions of the adenoviral genome for propagation, and does not requirecomplementation of any other deficiency of the adenoviral genome forpropagation.

The adenovirus or adenoviral vector, when deficient in multiplereplication-essential gene functions of the adenoviral genome (e.g., anE1/E4-deficient adenoviral vector), can include a spacer sequence toprovide viral growth in a complementing cell line similar to thatachieved by adenoviruses or adenoviral vectors deficient in a singlereplication-essential gene function (e.g., an E1-deficient adenoviralvector). The spacer sequence can contain any nucleotide sequence orsequences which are of a desired length, such as sequences at leastabout 15 base pairs (e.g., between about 15 nucleotides and about 12,000nucleotides), preferably about 100 nucleotides to about 10,000nucleotides, more preferably about 500 nucleotides to about 8,000nucleotides, even more preferably about 1,500 nucleotides to about 6,000nucleotides, and most preferably about 2,000 to about 3,000 nucleotidesin length, or a range defined by any two of the foregoing values. Thespacer sequence can be coding or non-coding and native or non-nativewith respect to the adenoviral genome, but does not restore thereplication-essential function to the deficient region. The spacer alsocan contain an expression cassette. More preferably, the spacercomprises a polyadenylation sequence and/or a gene that is non-nativewith respect to the adenovirus or adenoviral vector. The use of a spacerin an adenoviral vector is further described in, for example, U.S. Pat.No. 5,851,806 and International Patent Application Publication WO1997/021826.

By removing part of the adenoviral genome, for example, the E1, E3, andE4 regions of the adenoviral genome, the resulting adenovirus oradenoviral vector is able to accept inserts of exogenous nucleic acidsequences while retaining the ability to be packaged into adenoviralcapsids. An exogenous nucleic acid sequence can be inserted at anyposition in the adenoviral genome so long as insertion in the positionallows for the formation of adenovirus or the adenoviral vectorparticle. The exogenous nucleic acid sequence preferably is positionedin the E1 region, the E3 region, or the E4 region of the adenoviralgenome.

The replication-deficient adenovirus or adenoviral vector of theinvention can be produced in complementing cell lines that provide genefunctions not present in the replication-deficient adenovirus oradenoviral vector, but required for viral propagation, at appropriatelevels in order to generate high titers of viral vector stock. Suchcomplementing cell lines are known and include, but are not limited to,293 cells (described in, e.g., Graham et al., J. Gen. Virol., 36: 59-72(1977)), PER.C6 cells (described in, e.g., International PatentApplication Publication WO 1997/000326, and U.S. Pat. Nos. 5,994,128 and6,033,908), and 293-ORF6 cells (described in, e.g., International PatentApplication Publication WO 1995/034671 and Brough et al., J. Virol., 71:9206-9213 (1997)). Other suitable complementing cell lines to producethe replication-deficient adenovirus or adenoviral vector of theinvention include complementing cells that have been generated topropagate adenoviral vectors encoding transgenes whose expressioninhibits viral growth in host cells (see, e.g., U.S. Patent ApplicationPublication No. 2008/0233650). Additional suitable complementing cellsare described in, for example, U.S. Pat. Nos. 6,677,156 and 6,682,929,U.S. Patent Application Publication No. 2008/0233650 A1, andInternational Patent Application Publication WO 2003/020879. In someinstances, the cellular genome need not comprise nucleic acid sequences,the gene products of which complement for all of the deficiencies of areplication-deficient adenoviral vector. One or morereplication-essential gene functions lacking in a replication-deficientadenoviral vector can be supplied by a helper virus, e.g., an adenoviralvector that supplies in trans one or more essential gene functionsrequired for replication of the replication-deficient adenovirus oradenoviral vector. Alternatively, the inventive adenovirus or adenoviralvector can comprise a non-native replication-essential gene thatcomplements for the one or more replication-essential gene functionslacking in the inventive replication-deficient adenovirus or adenoviralvector. For example, an E1/E4-deficient adenoviral vector can beengineered to contain a nucleic acid sequence encoding E4 ORF 6 that isobtained or derived from a different adenovirus (e.g., an adenovirus ofa different serotype than the inventive adenovirus or adenoviral vector,or an adenovirus of a different species than the inventive adenovirus oradenoviral vector).

In addition to the nucleic acid encoding the one or more HSV antigens,the vector also can comprise gene(s) encoding one or moreimmunostimulatory/regulatory molecules, cytokines, or other moleculesthat can enhance an immune response to HSV. The nucleic acid, as well asany other exogenous gene(s), preferably are inserted into a site orregion (insertion region) in the vector that does not affect virusviability of the resultant recombinant virus. Such regions can bereadily identified by testing segments of virus DNA for regions thatallow recombinant formation without seriously affecting virus viabilityof the recombinant virus.

The vector preferably also comprises expression control sequences, suchas promoters, enhancers, polyadenylation signals, transcriptionterminators, internal ribosome entry sites (IRES), and the like, thatprovide for the expression of the nucleic acid sequence in a host cell.Exemplary expression control sequences are known in the art and aredescribed in, for example, Goeddel, Gene Expression Technology: Methodsin Enzymology, Vol. 185, Academic Press, San Diego, Calif. (1990).Ideally, the HSV antigen-encoding nucleic acid sequence is operablylinked to a promoter and a polyadenylation sequence. A large number ofpromoters, including constitutive, inducible, and repressible promoters,from a variety of different sources are well known in the art.Representative sources of promoters include for example, virus, mammal,insect, plant, yeast, and bacteria, and suitable promoters from thesesources are readily available, or can be made synthetically, based onsequences publicly available, for example, from depositories such as theATCC as well as other commercial or individual sources. Promoters can beunidirectional (i.e., initiate transcription in one direction) orbi-directional (i.e., initiate transcription in either a 3′ or 5′direction). Non-limiting examples of promoters include, for example, theT7 bacterial expression system, pBAD (araA) bacterial expression system,the cytomegalovirus (CMV) promoter, the SV40 promoter, and the RSVpromoter. Inducible promoters include, for example, the Tet system (U.S.Pat. Nos. 5,464,758 and 5,814,618), the Ecdysone inducible system (No etal., Proc. Natl. Acad. Sci., 93: 3346-3351 (1996)), the T-REx™ system(Invitrogen, Carlsbad, Calif.), LACSWITCH™ System (Stratagene, SanDiego, Calif.), and the Cre-ERT tamoxifen inducible recombinase system(Indra et al., Nuc. Acid. Res., 27: 4324-4327 (1999); Nuc. Acid. Res.,28: e99 (2000); U.S. Pat. No. 7,112,715; and Kramer & Fussenegger,Methods Mol. Biol., 308: 123-144 (2005)).

A promoter can be selected by matching its particular pattern ofactivity with the desired pattern and level of expression of anantigen(s) (e.g., the inventive or wild-type UL19 and/or UL47 antigens).For example, the vector can comprise two or more nucleic acid sequencesthat encode different antigens and are operably linked to differentpromoters displaying distinct expression profiles. In this regard, afirst promoter can be selected to mediate an initial peak of antigenproduction, thereby priming the immune system against an encodedantigen. A second promoter can be selected to drive production of thesame or different antigen such that expression peaks several days afterthat of the first promoter, thereby “boosting” the immune system againstthe antigen. Alternatively, a hybrid promoter can be constructed whichcombines the desirable aspects of multiple promoters. In that antigenscan be toxic to eukaryotic cells, it may be advantageous to modify thepromoter to decrease activity in complementing cell lines used topropagate the vector.

To optimize protein production, preferably the antigen-encoding nucleicacid sequence further comprises a polyadenylation site following thecoding sequence. Any suitable polyadenylation sequence can be used,including a synthetic optimized sequence, as well as the polyadenylationsequence of BGH (bovine growth hormone), polyoma virus, TK (thymidinekinase), EBV (Epstein Barr virus), and the papillomaviruses, includingHPV (human papillomavirus) and BPV (bovine papilloma virus). A preferredpolyadenylation sequence is the SV40 (Human Sarcoma Virus-40)polyadenylation sequence. Also, preferably all the proper transcriptionsignals (and translation signals, where appropriate) are correctlyarranged such that the nucleic acid sequence is properly expressed inthe cells into which it is introduced. If desired, the nucleic acidsequence also can incorporate splice sites (i.e., splice acceptor andsplice donor sites) to facilitate mRNA production.

In one embodiment, the HSV antigen-encoding nucleic acid sequencefurther comprises the appropriate sequences for processing, secretion,intracellular localization, and the like. The HSV antigen-encodingnucleic acid sequence can be operably linked to a signal sequence, whichtargets a protein to cellular machinery for secretion. Appropriatesignal sequences include, but are not limited to, leader sequences forimmunoglobulin heavy chains and cytokines (see, for example, Ladunga etal., Current Opinions in Biotechnology, 11: 13-18 (2000)). Other proteinmodifications can be required to secrete a protein from a host cell,which can be determined using routine laboratory techniques. Preparingexpression constructs encoding antigens and signal sequences is furtherdescribed in, for example, U.S. Pat. No. 6,500,641. Methods of secretingnon-secretable proteins are further described in, for example, U.S. Pat.No. 6,472,176 and International Patent Application Publication WO2002/048377.

An HSV antigen encoded by the nucleic acid sequence of the vector alsocan be modified to attach or incorporate the antigen on a host cellsurface. In this respect, the antigen can comprise a membrane anchor,such as a gpi-anchor, for conjugation onto a cell surface. Atransmembrane domain can be fused to the antigen to incorporate aterminus of the antigen protein into the cell membrane. Other strategiesfor displaying peptides on a cell surface are known in the art and areappropriate for use in the context of the invention.

One or more of the inventive nucleic acid sequences, vectors, orpolypeptides (alone or in further combination with other HSV antigensand/or other antigens), desirably in the form of a composition thatincludes a suitable carrier, can be administered to an animal,preferably a mammal, and most preferably a human. The human preferablyis in a population that has a high risk of acquiring HSV or already hasHSV.

Infection of an individual (e.g., human) with HSV can lead to HSVdisease, wherein the infected individual demonstrates symptoms such ascold sores, itching or tingling sensations in the genital or anal area,small fluid-filled blisters that can burst leaving small painful sores(genital blisters), painful urination (due to the passing of urine overthe open sores), headaches, backaches, and flu-like symptoms, includingswollen glands or fever. While not wishing to be bound by any particulartheory, the administration of the inventive nucleic acids, vectors,polypeptides, and compositions thereof leads to the treatment and/orprevention of HSV disease by reducing the presence of HSV virus and/orsymptoms in an individual.

The invention provides a method of inducing an immune response againstHSV in a mammal, a method of treating or preventing HSV disease in amammal, a method of inducing a T cell response against HSV in a mammal,a method of reducing HSV viral shedding in a mammal, and a method ofinducing an antibody response against HSV in a mammal. The HSV can beHSV-1 or HSV-2. The methods comprise administering to the mammal acomposition comprising one or more of the inventive nucleic acidsequences or inventive vectors and a pharmaceutically acceptablecarrier, whereupon the one or more nucleic acid sequences encoding theHSV antigen(s) and/or other antigens is expressed in the mammal toproduce the one or more HSV antigens and/or other antigens and therebyinduce an immune response against HSV in the mammal, treat or preventHSV disease in the mammal, induce a T cell response against HSV in themammal, and/or induce an antibody response against HSV in the mammal.

Alternatively, the methods comprise administering to the mammal acomposition comprising the inventive UL47 polypeptide (alone or infurther combination with the inventive and/or wild-type UL19 antigenand/or other antigens) and a pharmaceutically acceptable carrier tothereby induce an immune response against HSV in the mammal, treat orprevent HSV disease in the mammal, or induce a T cell response againstHSV in the mammal, and/or induce an antibody response against HSV in themammal.

The immune response can be a humoral immune response, a cell-mediatedimmune response, or a combination of humoral and cell-mediated immunity.Ideally, the immune response provides protection upon subsequentchallenge with HSV. However, protective immunity is not required in thecontext of the invention. The inventive method also can be used forantibody production.

The invention provides a composition comprising (a) one or more of theinventive nucleic acid sequences, one or more of the inventive vectors,or one or more of the inventive polypeptides (alone or in furthercombination with other HSV antigens and/or other antigens) and (b) acarrier (e.g., a pharmaceutically acceptable carrier). The compositiondesirably is a physiologically acceptable (e.g., pharmaceuticallyacceptable) composition, which comprises a carrier, preferably aphysiologically (e.g., pharmaceutically) acceptable carrier, and one ormore of the inventive nucleic acid sequences, vectors, or polypeptides(alone or in further combination with other HSV antigens and/or otherantigens). Any suitable carrier can be used within the context of theinvention, and such carriers are well known in the art. The choice ofcarrier will be determined, in part, by the particular use of thecomposition, e.g., administration to an animal and the particular methodused to administer the composition to the animal. The compositionoptionally can be sterile. Ideally, in embodiments in which the vectoris a replication-deficient adenoviral vector, the composition preferablyis free of replication-competent adenovirus (RCA) contamination (e.g.,the composition comprises less than about 1% of replication-competentadenovirus on the basis of the total adenoviruses in the composition).Most desirably, such a composition is RCA-free. Adenoviral vectorcompositions and stocks that are RCA-free are described in U.S. Pat. No.5,944,106 and International Patent Application Publication WO1995/034671.

To enhance the immune response generated against an HSV antigen, animmune stimulator, or a nucleic acid sequence that encodes an immunestimulator, also can be administered to the mammal (e.g., as a componentof the inventive compositions). Immune stimulators also are referred toin the art as “adjuvants,” and include, for example, cytokines,chemokines, or chaperones. Cytokines include, for example, MacrophageColony Stimulating Factor (e.g., GM-CSF), Interferon Alpha (IFN-α),Interferon Beta (IFN-β), Interferon Gamma (IFN-γ), interleukins (IL-1,IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12, IL-13, IL-15, IL-16,and IL-18), the tumor necrosis factor (TNF) family of proteins,Intercellular Adhesion Molecule-1 (ICAM-1), LymphocyteFunction-Associated antigen-3 (LFA-3), B7-1, B7-2, FMS-related tyrosinekinase 3 ligand, (Flt3L), vasoactive intestinal peptide (VIP), LIGHT(also known as TNFSFI4 or HVEM-L), and CD40 ligand. Chemokines include,for example, B Cell-Attracting chemokine-1 (BCA-1), Fractalkine,Melanoma Growth Stimulatory Activity protein (MGSA), Hemofiltrate CCchemokine 1 (HCC-1), Interleukin 8 (IL-8), Interferon-stimulated T cellalpha chemoattractant (I-TAC), Lymphotactin, Monocyte ChemotacticProtein 1 (MCP-1), Monocyte Chemotactic Protein 3 (MCP-3), MonocyteChemotactic Protein 4 (CP-4), Macrophage-Derived Chemokine (MDC), amacrophage inflammatory protein (MIP), Platelet Factor 4 (PF4),Regulated on Activation Normal T Cell Expressed and Secreted Chemokine(RANTES), Breast and Kidney-Expressed Chemokine (BRAK), eotaxin, exodus1-3, and the like. Chaperones include, for example, the heat shockproteins Hsp70, Hsc70, and Hsp40.

The composition also can comprise other antiviral drugs, such asnucleoside analogs, peptide analogs, and small molecules that targetviral transcription, translation, entry, and/or coating. For example,the composition can comprise acyclovir, foscarnet, ribavirin,interferons, or any combination thereof.

Suitable formulations for the composition include aqueous andnon-aqueous solutions, isotonic sterile solutions, which can containanti-oxidants, buffers, and bacteriostats, and aqueous and non-aqueoussterile suspensions that can include suspending agents, solubilizers,thickening agents, stabilizers, and preservatives. The formulations canbe presented in unit-dose or multi-dose sealed containers, such asampules and vials, and can be stored in a freeze-dried (lyophilized)condition requiring only the addition of the sterile liquid carrier, forexample, water, immediately prior to use. Extemporaneous solutions andsuspensions can be prepared from sterile powders, granules, and tablets.Preferably, the carrier is a buffered saline solution. If the vector isan adenoviral vector, then the composition preferably is formulated toprotect the adenoviral vector from damage prior to administration. Forexample, the composition can be formulated to reduce loss of theadenoviral vector on devices used to prepare, store, or administer theadenoviral vector, such as glassware, syringes, or needles. Thecomposition can be formulated to decrease the light sensitivity and/ortemperature sensitivity of the adenoviral vector. To this end, thecomposition preferably comprises a pharmaceutically acceptable liquidcarrier, such as, for example, those described above, and a stabilizingagent selected from the group consisting of polysorbate 80, L-arginine,polyvinylpyrrolidone, trehalose, and combinations thereof. Use of such acomposition will extend the shelf life of the vector, facilitateadministration, and increase the efficiency of the inventive method.Formulations for adenoviral vector-containing compositions are furtherdescribed in, for example, U.S. Pat. No. 6,225,289, U.S. Pat. No.6,514,943, and International Patent Application Publication WO2000/034444.

The composition also can be formulated to enhance transductionefficiency. In addition, the vector can be present in a composition withother therapeutic or biologically-active agents. For example, factorsthat control inflammation, such as ibuprofen or steroids, can be part ofthe composition to reduce swelling and inflammation associated with invivo administration of the vector. As discussed herein, immune systemstimulators or adjuvants, e.g., interleukins, lipopolysaccharide, ordouble-stranded RNA, can be administered to enhance or modify any immuneresponse to the HSV antigen. Antibiotics, i.e., microbicides andfungicides, can be present to treat existing infection and/or reduce therisk of future infection, such as infection associated with genetransfer procedures.

Any route of administration can be used to deliver the composition tothe mammal. Indeed, although more than one route can be used toadminister the composition, a particular route can provide a moreimmediate and more effective reaction than another route. Preferably,the composition is administered via intramuscular injection orintradermal, subcutaneous, oral, colorectal, or intranasaladministration. The composition also can be applied or instilled intobody cavities (e.g., intravaginal), absorbed through the skin (e.g., viaa transdermal patch), inhaled, ingested, topically applied to tissue, oradministered parenterally via, for instance, intravenous, peritoneal, orintraarterial administration. For example, when parenteraladministration is employed, the composition can be administered in thearm (e.g., upper arm), buttocks, thigh, and/or face (e.g., in and aroundthe lips). The administration can be via standard injection devices(e.g., needle and syringe), via a mechanical device or device usingelectricity, heat, cold, beads, or magnetic waves, and/or via microneedles or dissolving maltose-based needles (see, e.g., Eisenstein M.,Nature Biotechnology, 29(2): 107-109 (2011).

The composition can be administered in or on a device that allowscontrolled or sustained release, such as a sponge, biocompatiblemeshwork, mechanical reservoir, or mechanical implant. Implants (see,e.g., U.S. Pat. No. 5,443,505), devices (see, e.g., U.S. Pat. No.4,863,457), such as an implantable device, e.g., a mechanical reservoiror an implant or a device comprised of a polymeric composition, areparticularly useful for administration of the composition. Thecomposition also can be administered in the form of sustained-releaseformulations (see, e.g., U.S. Pat. No. 5,378,475) comprising, forexample, gel foam, hyaluronic acid, gelatin, chondroitin sulfate, apolyphosphoester, such as bis-2-hydroxyethyl-terephthalate (BHET),and/or a polylactic-glycolic acid.

The dose of the inventive nucleic acids, inventive vectors, and/orwild-type or inventive polypeptides as described herein administered tothe mammal will depend on a number of factors, including the extent ofany side-effects, the particular route of administration, and the like.The dose ideally comprises an “effective amount” of the inventivenucleic acids, inventive vectors, and/or wild-type or inventivepolypeptides as described herein. For example, the dose of an inventivevector desirably is a dose of vector which provokes a desired immuneresponse in the mammal. A single dose of vector (e.g., adenoviralvector) desirably comprises at least about 1×10⁵ particles (which alsois referred to as particle units) of vector. The dose preferably is atleast about 1×10⁶ particles (e.g., about 1×10⁶-1×10¹² particles), morepreferably at least about 1×10⁷ particles, more preferably at leastabout 1×10⁸ particles (e.g., about 1×10⁸-1×10¹¹ particles or about1×10⁸-1×10¹² particles), and most preferably at least about 1×10⁹particles (e.g., about 1×10⁹-1×10¹⁰ particles or about 1×10⁹-1×10¹²particles), or even at least about 1×10¹⁰ particles (e.g., about1×10¹⁰-1×10¹² particles) of the vector. The dose desirably comprises nomore than about 1×10¹⁴ particles, preferably no more than about 1×10¹³particles, even more preferably no more than about 1×10¹² particles,even more preferably no more than about 1×10¹¹ particles, and mostpreferably no more than about 1×10¹⁰ particles of the vector. In otherwords, a single dose of the vector (especially adenoviral vector) cancomprise, for example, about 1×10⁶ particle units (PU), 2×10⁶ PU, 4×10⁶PU, 1×10⁷ PU, 2×10⁷ PU, 4×10⁷ PU, 1×10⁸ PU, 2×10⁸ PU, 4×10⁸ PU, 1×10⁹PU, 2×10⁹ PU, 4×10⁹ PU, 1×10¹⁰ PU, 2×10¹⁰ PU, 4×10¹⁰ PU, 1×10¹¹ PU,2×10¹¹ PU, 4×10¹¹ PU, 1×10¹² PU, 2×10¹² PU, or 4×10¹² PU of the vector.

Administration of the composition containing the inventive vector can beone component of a multistep regimen for inducing an immune responseagainst HSV in a mammal, treating or preventing HSV disease in themammal, inducing a T cell response against HSV in the mammal, and/orinducing an antibody response against HSV in the mammal. In thisrespect, the method further comprises administering to the mammal aboosting composition after administering the vector to the mammal. Inthis embodiment, therefore, the immune response is “primed” uponadministration of the composition containing the first vector (orcomposition of multiple vectors), and is “boosted” upon administrationof the boosting composition. Alternatively, the inventive method furthercomprises administering to the mammal a priming composition to themammal prior to administering the vector to the mammal. In thisembodiment, therefore, the immune response is “primed” uponadministration of the priming composition, and is “boosted” uponadministration of the composition containing the vector. The inventivemethod can comprise multiple administrations of the same entity ordifferent entities (e.g., two, three, four, five, or six times or more).

Each of the priming composition and the boosting composition desirablycomprises one or more vectors that comprise a nucleic acid sequenceencoding one or more HSV antigens. Any suitable vector can be employed,including viral and non-viral vectors, as described herein. Examples ofsuitable viral vectors include, but are not limited to, retroviralvectors, adeno-associated virus vectors, vaccinia virus vectors,herpesvirus vectors, parainfluenza-RSV chimeric vectors (PIV-RSV), andadenoviral vectors. Examples of suitable non-viral vectors include, butare not limited to, plasmids, liposomes, nanoparticles, and molecularconjugates (e.g., transferrin). Preferably, the priming composition orthe boosting composition is a plasmid or an adenoviral vector.Alternatively, an immune response can be primed or boosted byadministration of an HSV protein itself (e.g., an antigenic HSV protein)with or without a suitable adjuvant (e.g., alum, QS-21, insulin-derivedadjuvant, etc.), a live-attenuated HSV particle, a virus-like particle,and the like. When the priming composition and/or the boostingcomposition is an adenoviral vector, it can be an adenoviral vectorderived from any human or non-human animal as described herein.

The priming and boosting compositions can be the same or different. Whenthe priming and the boosting compositions are the same, the compositionsare deemed “homologous.” When the priming and the boosting compositionsare different, the compositions are deemed “heterologous.”

In one embodiment, the vectors used in the priming and boostingcompositions can comprise any combination of the following, wherein thepriming and boosting compositions can be the same or different: humanserotype 28 adenoviral (Ad28) vectors, Ad28 vectors with hexon from adifferent adenoviral serotype (Ad28 H), Ad28 with hexon and fiber (e.g.,knob) from a different adenoviral serotype (Ad28 H/F), human serotype 5adenoviral (Ad5) vectors, Ad5 with hexon from a different adenoviralserotype (Ad5 H), Ad5 with hexon and fiber from a different adenoviralserotype (Ad5 H/F), and other adenoviral vectors (e.g., gorillaadenoviral vectors) as described herein.

In a preferred embodiment, the priming composition and/or the boostingcomposition comprises a human adenoviral vector (e.g., serotype 5, 28,or 35) or another adenoviral vector, including but not limited to otherspecies. For example, a priming composition containing a human serotype28 adenoviral vector can be administered to a human, followed byadministration of a boosting composition containing another adenoviralvector (e.g., adenoviral vector isolated from gorilla). Alternatively, apriming composition containing a human serotype 28 adenoviral vector canbe administered to a human, followed by administration of a boostingcomposition containing a modified human serotype 28 adenoviral vectorcomprising hexon and/or fiber (e.g., knob) from a different serotype(e.g., serotype 45) adenoviral vector. In another embodiment, a primingcomposition containing another species of adenoviral vector (e.g.,adenoviral vector isolated from gorilla) can be administered to a human,followed by a second administration of the same composition. Theinvention encompasses the use of any combination of human (e.g., Ad28,Ad28 H, Ad28 H/F, Ad5, Ad5 H, and/or Ad5 H/F) and/or other adenoviralvectors (e.g., adenoviral vectors isolated from gorilla) encoding one ormore HSV antigens and/or other antigens in the priming or boostingcomposition.

When two or more administrations are employed for the prime/boost, thevectors of the priming composition and the boosting composition(s)(which vectors can be the same or different) desirably comprise the sameexogenous nucleic acid sequence(s) (e.g., at least one nucleic acidsequence encoding the same HSV antigen or multiple (i.e., two or more)nucleic acid sequences encoding the same HSV antigen or antigens). Inanother embodiment, the vectors of the priming composition and/or theboosting composition(s) (which vectors can be the same or different) cancomprise different exogenous nucleic acid sequences encoding one, two,or more different HSV antigens. The priming and/or boostingcomposition(s) also can contain one or more cytokines or adjuvants.

Administration of the priming composition and the boosting compositioncan be separated by any suitable timeframe (e.g., at least about 1 week,2 weeks, 3 weeks, 4 weeks, 8 weeks, 12 weeks, 16 weeks, 24 weeks, 52weeks, or a range defined by any two of the foregoing values). Theboosting composition preferably is administered to a mammal (e.g., ahuman) at least about 1 week (e.g., 2 weeks, 3 weeks, 4 weeks, 5 weeks,6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13weeks, 14 weeks, 15 weeks, 16 weeks, 20 weeks, 24 weeks, 28 weeks, 35weeks, 40 weeks, 50 weeks, 52 weeks, or a range defined by any two ofthe foregoing values) following administration of the primingcomposition. More than one dose of priming composition and/or boostingcomposition can be provided in any suitable timeframe. The dose of thepriming composition and boosting composition administered to the mammaldepends on a number of factors, including the extent of anyside-effects, the particular route of administration, and the like.

In preferred embodiments, the invention provides a method of inducing animmune response against HSV in a mammal, a method of treating orpreventing HSV disease in a mammal, a method of inducing a T cellresponse against HSV in a mammal, and a method of inducing an antibodyresponse against HSV in a mammal, which method comprises (a)administering to the mammal a priming composition comprising one or morereplication-deficient adenoviral vectors comprising (i) one or more ofthe inventive nucleic acid sequences and/or (ii) one or more nucleicacid sequences comprising the wild-type UL19 and/or UL47 sequence(s) anda pharmaceutically acceptable carrier, and (b) administering to themammal a boosting composition comprising (i) one or more of theinventive nucleic acid sequences and/or (ii) one or more nucleic acidsequences comprising the wild-type UL19 and/or UL47 sequence(s) and apharmaceutically acceptable carrier. The administration of the primingcomposition and the boosting composition can be separated by anysuitable length of time as described herein, but preferably is at leastabout 1 week. The administration of the initial administration andsubsequent administrations can be separated by any suitable length oftime as described herein, but preferably is at least about 1 week apart.The administration of the boosting composition, or subsequentadministrations, desirably induces an enhanced immune response (e.g. Tcell response), as compared to the immune response induced after theadministration of the priming composition, or initial administration,alone.

The inventive methods also can include the simultaneous, subsequent,and/or sequential administration of other antiviral drugs, such asnucleoside analogs, peptide analogs, and small molecules that targetviral transcription, translation, entry, and/or coating, such asacyclovir, foscarnet, ribavirin, interferons, or any combinationthereof.

The following examples further illustrate the invention but, of course,should not be construed as in any way limiting its scope.

EXAMPLE 1

This example demonstrates the construction of an adenovirus comprisingthe inventive nucleic acid sequence encoding an HSV antigen.

A human serotype 28 adenoviral vector was prepared with a deletion inthe E1 region, the replacement of the hexon region with the hexon regionof a human serotype 45 adenovirus, and the insertion of the nucleic acidsequence comprising SEQ ID NO: 5 (Ad28UL19 H(Ad45)). Ad28UL19 H(Ad45) isreplication-deficient due to the deletion of the essential functionprovided by E1. The CMV promoter and transgene expression cassettecomprising SEQ ID NO: 5 were introduced in place of the E1 sequences.The expression cassette, located at the E1 region deletion junction, isright-to-left with respect to the viral genome.

Ad28UL19 H(Ad45) has an E1 region deletion of bases 462 through 3111 ofthe Ad28 genomic sequence. The deletion includes Ad28E1A and part of theE1B early proteins, which renders the vector replication incompetent innoncomplementing cell lines.

A description of the vector construction is as follows. The plasmidpAC28E1(t.UL19)H(45) was constructed that encodes the entireAd28UL19H(Ad45) adenoviral vector genome with the CMV.UL19 expressioncassette in the E1 region and the Ad45 hexon in place of the Ad28 hexon.A single genetic clone of the final vector genome was achieved by twosequential colony-growth steps in bacteria. This viral vector genomeencoded by the plasmid was converted to a viral vector upon introductioninto mammalian cells that complement for adenoviral vector growth.Subsequent expansion via serial passaging was performed to generateadenoviral vector stocks.

A similar method was used to incorporate the Ad45 fiber (shaft withknob) domain in place of the Ad28 fiber. This entire method was repeatedfor other HSV2 antigens cloned into the adenoviral vector base, Ad28.

EXAMPLE 2

This example describes animal models for the evaluation of antiviralefficacy.

Mouse Model of Primary HSV-1/HSV-2 Challenge:

The primary screening model provides a rapid initial evaluation ofantiviral efficacy against HSV primary infection with both clinical andvirological endpoints. This model utilizes intravaginal inoculation offemale mice (25 g) with HSV-1 or HSV-2 to evaluate potential antiviraltherapies as well as vaccine/adjuvant candidates. Animals are followeddaily for signs and systems of herpes disease, and vaginal swabs areobtained to evaluate the effect of therapy on viral replication. Singleor combined antiviral therapies can be administered by any suitablemeans, including, but not limited to, topically, orally or systemically,and can be given at varying intervals begun before or after viralchallenge. Dose range studies also can be carried out.

Dose and route of administration are individualized for eachexperimental agent. However, for each intravaginal dose, 15-30 μl ofproduct typically is required. Treatment group size is typically 12-16mice.

Ovariectomized Mouse Model of HSV-2 Challenge:

Four week old BALB/c mice are ovariectomized (OVEX), allowed to recoverfor two weeks, and randomly placed into groups of twelve animals forexperimental groups. Mice are infected intra-vaginally with PBS or adose from 10 plaque forming units (PFU)-100000 PFU of HSV2, Strain 186,and monitored daily for survival out to Day 21 post-HSV2 infection.Vaginal swabs are collected on Day −1, +2, +6, and +7. Using thismethod, HSV2 infection in ovariectomized female BALB/c mice has beenshown to exhibit dose dependent survival, as illustrated in FIG. 7.

Guinea Pig Model of Primary Genital HSV-2 Infection:

Because genital herpes disease in the guinea pig more closely resembleshuman disease, guinea pigs are used as a second species for therapieswith demonstrated efficacy against HSV in mice. As with humans, genitalHSV infection in guinea pigs is a self limited vesiculoulcerativedisease which is followed by healing, the establishment of latency, andthen both spontaneous and inducible symptomatic and asymptomaticrecurrences. The model utilizes intravaginal inoculation of femaleHartley guinea pigs and provides both clinical and virologic indices toassess both the effect of treatment on primary disease as well as on thefrequency or severity of subsequent recurrent infections. Antiviraltherapy can be administered by any suitable means including orally,topically, or systemically and can be given at varying intervalsbeginning before or after virus challenge. Following intravaginalinoculation, animals are followed daily for the development of genitalherpes using a validated genital herpes scoring system. Vaginal swabsare also obtained to evaluate the effect against viral replication.Because this is a non lethal model, animals can be sacrificed at theconclusion of the experiment to evaluate the effects of treatment onlatency. This model can be adapted to evaluate antiviral activityagainst available drug resistant strains (acyclovir (ACV) andfoscarnet).

Dose and route of administration and duration of treatment areindividualized for each experimental agent. For assessing drugrequirements the average weight of the animals typically is 300 g.Treatment group size is typically 10-15 animals.

Guinea Pig Model of Recurrent Genital HSV-2 Infection:

The guinea pig model of genital herpes is unique in that, after recoveryfrom primary genital infection, animals experience spontaneous recurrentgenital lesions as well as viral shedding in the absence of lesions.This allows a candidate compound to be evaluated for efficacy incontrolling recurrent disease. Female Hartley guinea pigs who haverecovered from symptomatic primary genital infection are randomized intotreatment groups for antiviral treatments beginning on day 15post-infection (PI) and continuing for 48 days thereafter. Treatmentscan be administered by any suitable means including orally, topically,or systemically. The indices for these studies include quantificationand severity assessment of recurrent episodes during treatment and for21 days following cessation of treatment. Additionally, vaginal swabsare collected to evaluate any impact on shedding.

Dose and route of administration are individualized for eachexperimental agent. Treatment group size is typically 10-15 animals.

Model of Neonatal HSV-2 Infection in Guinea Pigs:

The model of neonatal HSV infection mimics the natural history ofinfection in the human newborn. This model is available to evaluatecandidate antiviral drug therapies and combined therapeutic approachesincluding combinations of antivirals or antivirals and immunemodulators. Additionally, this model can be used to evaluate theefficacy of candidate vaccines by measuring the protection afforded by atransplacental antibody. In this model newborn, Hartley guinea pigs areinoculated intranasally with HSV-2 within 48 hours of delivery. Newbornanimals are then randomized to receive experimental drug, placebo, orACV (control). Animals are evaluated daily for evidence of cutaneousherpetic disease and weight gain as well as pulmonary disorders, CNSsymptoms, and death. Surviving animals are followed for 45 days toassess the effectiveness of therapy on the incidence and frequency ofcutaneous herpetic recurrences.

Dose and route of administration are individualized for eachexperimental agent. Duration of treatment is typically 10 days or more.A positive control of ACV (60 mg/kg/day) twice daily (i.e., BID) can beused. Newborn guinea pigs typically weigh about 60-100 gms.

Additional information regarding animal models for use with theinvention can be found in Hoshino et al., J. Virol., 79(1): 410-418(2005).

EXAMPLE 3

This example demonstrates that the administration of a vector comprisingone or more nucleic acid sequences encoding one or more HSV antigensinduces an immune response (T cell response) against HSV.

Two modified serotype 28 adenoviral vectors comprising hexon and fiberfrom a serotype 45 adenoviral vector (Ad28 H/F) were produced using themethods described in Example 1. The first vector comprised a nucleicacid sequence comprising SEQ ID NO: 7 (a wild-type (non-modified) UL47;designated LW01), while the second vector comprised a nucleic acidsequence comprising SEQ ID NO: 5 (inventive UL19 nucleic acid sequence;designated LW02).

T cell response in mice following a single intramuscular administrationof each adenoviral vector-delivered antigen (1×10⁹ PU) was compared tonatural infection (1×10⁶ PFU of HSV administered intravaginally) asdescribed in Example 2. Animals infected with HSV showed symptoms of HSVinfection. As a negative control, animals were injected with finalformulation buffer (FFB-vehicle), which did not result in any inductionof HSV specific T cells. The results of this experiment are depicted inFIGS. 1A and 1B.

As demonstrated by the data set forth in FIGS. 1A and 1B, singleadministration of each of the adenoviral-delivered HSV antigens (thewild-type sequence of SEQ ID NO: 7 (LW01) and the inventive SEQ ID NO: 5(LW02)) resulted in a strong T cell response when compared to the T cellresponse generated by natural infection with a wild-type HSV-2 and the Tcell response of FFB-vaccinated animals.

The experiment was repeated with Ad28 H/F comprising a nucleic acidsequence comprising SEQ ID NO: 1 (inventive UL47; designated LW11). Theresults of this experiment are depicted in FIG. 2. As compared to Ad28H/F comprising the wild-type sequence of SEQ ID NO: 7 (LW01) and Ad28H/Fcomprising SEQ ID NO: 3 (inventive UL47 containing ubiquitin; designatedLW21), Ad28 H/F comprising LW11 resulted in a greater T cell response(see FIG. 2).

These results demonstrate that the administration of a vector (e.g., aserotype 28 adenoviral vector) comprising the inventive nucleic acidsequence encoding an HSV antigen induces an immune response (i.e.,increases T cell response) against HSV. In particular, vaccination witha vector comprising an inventive UL47 nucleic acid sequence inducedsuperior HSV-specific T cell levels as compared to a vector comprising awild-type UL47 nucleic acid sequence, whereas additional modificationsto the inventive UL47 nucleic acid sequence to include the ubiquitinsequence reduced HSV-specific T cell levels.

EXAMPLE 4

This example demonstrates the double administration (prime/boost) of avector comprising the inventive nucleic acid sequence encoding an HSVantigen.

Mice were intramuscularly administered a priming dose of 1×10⁹ PU of aserotype 28 adenoviral vector (Ad28) comprising SEQ ID NO: 7 (i.e., awild-type UL47 nucleic acid sequence) followed four weeks later by aboosting dose of 1×10⁹ PU of (i) Ad28 comprising SEQ ID NO: 7, (ii)modified serotype 28 adenoviral vector with hexon from serotype 45adenovirus (Ad28H) comprising SEQ ID NO: 7, (iii) modified serotype 28adenoviral vector with hexon and fiber from serotype 45 adenovirus(Ad28H/F) comprising SEQ ID NO: 7, or (iv) FFB (vehicle). The percentageof T cells was identified for each different boosting dose, and theresulting data is depicted in FIG. 3.

As demonstrated by the data set forth in FIG. 3, a heterologousprime/boost (Ad28 followed by Ad28 H or Ad28H/F) resulted in greaterHSV-specific T cell responses than homologous prime/boost (Ad28 followedby Ad28) or single administration (administration of Ad28 followed byvehicle).

This example demonstrates the efficacy of a multi-administration (e.g.,prime/boost) protocol with the inventive nucleic acids, vectors, andcompositions.

EXAMPLE 5

This example further demonstrates that the inventive nucleic acidsequence encoding an HSV antigen stimulates robust T cell responsesagainst the antigen.

Three different adenoviral vectors designated GC44 (SEQ ID NO: 10),GC45, (SEQ ID NO: 14), and GC46 (SEQ ID NO: 26) were modified by geneticengineering to (1) be replication-deficient by deletion of the E1 regionand (2) comprise SEQ ID NO: 5 (inventive UL19 nucleic acid sequence;designated LW02). More specifically, a nucleic acid sequence comprisingSEQ ID NO: 5 was introduced between a CMV promoter and the SV40 earlypolyA. The CMV promoter combines the CMV immediate early high expressionenhancer/promoter with tetracycline operator sites. Within this sequenceis the viral enhancer, CAAT box, TATA box, two copies of the 20nucleotide tetracycline operator sequence (tetO) from transposon Tn10,and the CMV transcription start site. The tetO sites are inactive inmammalian cells since tetracycline-based gene expression regulation isspecific for a prokaryotic system (see, e.g., Blau et al., Proc. Natl.Acad. Sci USA., 96(3): 797-799 (1999)). The tetO sites inhibit transgeneexpression when the viral vector is propagated in a cell line in thepresence of the tetracycline repressor. To further optimize theexpression of the nucleic acid sequence comprising SEQ ID NO: 5 from theCMV promoter, an artificial intron was created in the sequence byplacing a splice donor and a splice acceptor sequence upstream of theinitiation codon for the nucleic acid sequence comprising SEQ ID NO: 5.

The resulting adenoviral vectors designated GC44 LW02, GC45 LW02, andGC46 LW02 were propagated in a genetically stable 293-ORF6-cell linethat constitutively expresses the tetracycline repressor protein (TetR),which has been named M2A. The M2A cell line has been shown toefficiently reduce adenoviral vector transgene expression duringadenoviral vector construction and growth (see, e.g., U.S. PatentApplication Publication 2008/0233650 A1).

T cell response following single intramuscular administration of each ofthe adenoviral vectors comprising SEQ ID NO: 5 (1×10⁹ PU) was assessedin a mouse and compared to administration of a human adenoviral vectorserotype 5 (Ad5) comprising SEQ ID NO: 5 or control (FFB). The resultingdata is set forth in FIG. 4. As demonstrated by the data set forth inFIG. 4, single administration of each adenoviral vector comprising SEQID NO: 5 resulted in a strong T cell response.

These results demonstrate that the inventive nucleic acid sequenceencoding an HSV antigen stimulated a robust T cell response against theantigen. In particular, when delivered with GC44, GC45, and GC46, theinventive nucleic acid sequences resulted in high T cell response.

EXAMPLE 6

This example demonstrates that administration of a vector comprising aninventive nucleic acid sequence encoding an HSV antigen decreases HSVinfection symptoms (mean lesion score) and increases longevity(survival).

Ovariectomized female Balb/c mice were given a single intramuscularvaccination of 10⁸ pu of GC45 LW02. As a positive control, one group ofanimals was injected with FFB (vehicle). Wild-type HSV-2 wasadministered intravaginally two weeks subsequent to immunization. Onegroup of animals was not immunized but given Phosphate Buffered Saline(PBS) intravaginally as negative control for HSV infection. The animalswere evaluated for lesions and death rate, and the resulting data is setforth in FIGS. 5A and 5B.

As shown in FIG. 5A, a single administration with GC45 LW02 resulted ina decrease in mean lesion score for those animals as compared to theFFB-treated animals also receiving HSV infection. The PBS-treated(non-HSV infected) mice had no lesions as expected.

FIG. 5B shows that immunization with GC45 LW02 resulted in a highernumber of animals living both longer and not succumbing to the HSV-2disease as compared to the FFB-treated group. All the HSV-infectedanimals administered FFB succumbed to disease by Day 12. All of thePBS-treated (non-HSV infected) mice survived as expected.

EXAMPLE 7

This example demonstrates that a single administration of an adenoviralvector comprising an inventive nucleic acid sequence encoding an HSVantigen induces robust antigen-specific T cell responses.

A human serotype 28 adenoviral vector was prepared with a deletion inthe E1 region, the replacement of the hexon region and the fiber regionwith the hexon and fiber regions of a human serotype 45 adenovirus, andthe insertion of the inventive UL19-encoding nucleic acid sequencecomprising SEQ ID NO: 5 (Ad28HF.UL19), as described in Example 1.Gorilla adenoviral vectors designated GC46 (SEQ ID NO: 26) were preparedcontaining a deletion in the E1 region and an insertion of the inventiveUL19-encoding nucleic acid sequence comprising SEQ ID NO: 5 or theinventive UL47-encoding nucleic acid sequence comprising SEQ ID NO: 1,as described in Example 5. The resulting gorilla adenoviral vectors weredesignated GC46.UL19 and GC46.UL47.

Five BALB/c animals received a single intramuscular (IM) administration(1×10⁹ particle units (pu)) of GC46.UL19, GC46.UL47, or vehicle control.Another group of five BALB/c mice were either immunized with a single IMadministration (1×10⁹ pu) of Ad28HF.UL19, GC46.UL19, GC46.UL47, orvehicle control, or were infected with 1×10⁴ to 1×10⁶ plaque formingunits (PFU) of HSV2, Strain 186 intra-vaginally. Two weeks later,splenocytes were harvested and re-stimulated for 6 hours with UL19 andUL47-specific peptides for production of IFN-γ cytokine as measured byintracellular cytokine staining via FACS analysis. The results of theseexperiments are set forth in FIGS. 6A and 6B. A single administration ofeither Ad28HF.UL19, GC46.UL19, GC46.UL47 induced robust antigen-specificT cell responses that were significantly greater than T cell responsesinduced by HSV2 infection. One animal infected with HSV2 died frominfection before harvest. No T cell response was detected in animalsinfected with up to 1×10⁶ PFU of HSV2.

The results of this example demonstrate that a single administration ofthe inventive adenoviral vectors can induce strong HSV2-specific immuneresponses in animals.

EXAMPLE 8

This example demonstrates that a single administration of a blend ofadenoviral vectors encoding the inventive HSV2 antigens reduces HSV2symptoms in a mouse challenge model.

Ovariectomized BALB/c mice described in Example 2 were randomized intogroups of 12 animals. Mice were immunized IM with 1×10⁹ pu each of theadenoviral vector GC46.UL19, the adenoviral vector GC46.UL47, a blend ofGC46.UL19 and GC46.UL47 (1×10⁹ pu each), or PBS. Two weeks later, micewere challenged intra-vaginally with HSV2, Strain 186, and monitoreddaily for clinical symptoms. The results of this experiment are setforth in FIG. 8.

HSV2 viral shedding in treated mice was assayed using quantitative PCR(qPCR) and a plaque assay. Specifically, vaginal swabs were collectedfrom the experimental animals, and analyzed for (a) the amount of genomecopies detectable by quantitative PCR (qPCR) (LOD/LOQ=2/20) and (b) theamount of live virus titer by plaque assay on Vero cell monolayers(LOD/LOQ=1/10). The results of the qPCR and plaque assays are shown inFIGS. 9A and 9B.

The results of this example demonstrate that a single administration ofa blend of the inventive adenoviral vectors reduces symptoms of HSV2infection and viral shedding upon challenge with HSV2 in a mouse model.

EXAMPLE 9

This example demonstrates that a single administration of a blend ofadenoviral vectors encoding the inventive HSV2 antigens reduces theincidence and severity of HSV2 symptoms in a guinea pig model.

The guinea pig model of recurrent genital HSV-2 infection described inExample 2 was utilized for these experiments. Specifically, four to sixweek old female Hartley guinea pigs were infected intra-vaginally with5000 PFU of HSV2, Strain G, and randomized into treatment groups. On day9 following infection, animals received a single IM injection of 2×10⁸pu each of the adenoviral vector GC46.UL19 and the adenoviral vectorGC46.UL47 (described in Example 7). Animals not receiving any treatmentserved as a control. Daily observations for lesion incidence and lesionscores were performed. The results of these experiments are shown inFIGS. 10A and 10B. A single administration of a blend of GC46.UL19 andGC46.UL47 reduced the incidence and severity of HSV2 symptoms, and theseeffects were observed out to 63 days post immunization.

The results of this example demonstrate that a single administration ofa blend of the inventive adenoviral vectors reduces the recurrence andseverity of HSV2 lesions in a guinea pig model.

EXAMPLE 10

This example demonstrates that a homologous prime/boost immunizationmethod using the inventive adenoviral results in enhanced vaccineefficiency.

BALB/c mice were immunized with a single 1×10⁹ pu dose of GC46.UL19(described in Example 7), GC45.UL19 (referred to as “GC45 LW02” inExample 5), Ad28HF UL19 (described in Example 7), or vehicle control onDay 0. After 12 weeks, animals were immunized with a second dose of1×10⁹ pu of the same vector administered on day 0 (i.e., mice receivingGC46.UL19 on day 0 received a second dose of GC46.UL19 at 12 weeks). Twoweeks later, splenocytes were collected and re-stimulated for 6 hourswith UL19-specific peptides, and cytokine production was measured byintracellular staining via FACS. The results of this experiment areshown in FIG. 11. Repeat vaccination of mice with GC46 or GC45 vectorsresulted in a boost of vaccine efficacy which was not observed with theAd28HF vector.

The results of this example demonstrate that repeated administration ofgorilla adenoviral vectors encoding the inventive HSV2 UL19 antigenenhances HSV2 T cell responses as compared to repeated administration ofa human adenoviral vector encoding the same antigen.

EXAMPLE 11

This example demonstrates that a homologous prime/boost immunizationmethod using an adenoviral vector encoding the inventive UL19 and UL47antigens results in enhanced vaccine efficiency.

Using the methods described in Example 5, the GC46 adenoviral vectorGC46 UL19/UL47 was generated by genetic engineering to (1) bereplication-deficient by deletion of the E1 region and the E4 region,(2) comprise SEQ ID NO: 5 (inventive UL19 nucleic acid sequence), and(3) comprise SEQ ID NO: 1 (inventive UL47 nucleic acid sequence).

BALB/c mice were immunized IM on Day 0 with 1×10⁷ pu of the GC46UL19/UL47 vector or vehicle control. After one month (day 28), animalswere immunized with a second dose of 1×10⁷ pu of GC46 UL19/UL47, orvehicle control. One group of animals was not administered a boostingimmunization. Two weeks later, splenocytes and mucosa were collected andre-stimulated for 6 hours with UL19 and UL47-specific peptides, andcytokine production was measured by intracellular staining via FACS. Theresults of this experiment are shown in FIGS. 12A and 12B. Repeatedvaccination of mice with the GC46 UL19/UL47 vector resulted in a boostof vaccine efficacy with respect to both UL19-specific and UL47-specificimmune responses. A single IM administration of the GC46 UL19/UL47elicited mucosal T cell responses, and these mucosal T cell responseswere enhanced with repeated vaccination with the GC46 UL19/UL47 vector.

The results of this example demonstrate that a single IM administrationof a gorilla adenoviral vector encoding the inventive HSV2 UL19 antigenand the inventive HSV2 UL47 antigen elicit mucosal immunity in vivo, andthat both mucosal and systemic immune responses are enhanced withrepeated administration of the same vector.

EXAMPLE 12

This example demonstrates that a heterologous prime/boost immunizationmethod using the inventive adenoviral results in enhanced vaccineefficiency.

BALB/c mice were immunized on Day 0 with 1×10⁹ pu of the GC45.UL19adenoviral vector (referred to as “GC45 LW02” in Example 5), or vehiclecontrol. After three months (week 12), animals were immunized with 1×10⁹pu of GC45.UL19, 1×10⁹ pu of the GC46.UL19 vector (described in Example7), or vehicle control. Two weeks later, splenocytes were collected andre-stimulated for 6 hours with UL19- and UL47-specific peptides, andcytokine production was measured by intracellular staining via FACS. Theresults of this experiment are shown in FIG. 13. Administration ofGC45.UL19 as a prime followed by GC46.UL19 as a boost produced enhancedT cell responses as compared to administration of either GC45.UL19 orGC46.UL19 alone.

EXAMPLE 13

This example demonstrates that a single administration of an adenoviralvector comprising an inventive nucleic acid sequence encoding an HSVantigen induces durable T cell responses.

BALB/c mice were immunized on Day 0 with a single IM injection ofGC46.UL19 (described in Example 7). Mice were sacrificed at weeks 2, 5,14, and 26 following immunization. Splenocytes were harvested, andre-stimulated for 6 hours with UL19-specific peptides for production ofIFN-γ cytokine as measured by intracellular cytokine staining via FACSanalysis. The results of this experiment are shown in FIG. 14. Thepattern of CD8+ T cell responses induced following a singleadministration of GC46.UL19 were long-lasting (i.e., durable). Incontrast the T cell response is extremely low or non-existent followingnormal infection by HSV2.

All references, including publications, patent applications, andpatents, cited herein are hereby incorporated by reference to the sameextent as if each reference were individually and specifically indicatedto be incorporated by reference and were set forth in its entiretyherein.

The use of the terms “a” and “an” and “the” and similar referents in thecontext of describing the invention (especially in the context of thefollowing claims) are to be construed to cover both the singular and theplural, unless otherwise indicated herein or clearly contradicted bycontext. The terms “comprising,” “having,” “including,” and “containing”are to be construed as open-ended terms (i.e., meaning “including, butnot limited to,”) unless otherwise noted. Recitation of ranges of valuesherein are merely intended to serve as a shorthand method of referringindividually to each separate value falling within the range, unlessotherwise indicated herein, and each separate value is incorporated intothe specification as if it were individually recited herein. All methodsdescribed herein can be performed in any suitable order unless otherwiseindicated herein or otherwise clearly contradicted by context. The useof any and all examples, or exemplary language (e.g., “such as”)provided herein, is intended merely to better illuminate the inventionand does not pose a limitation on the scope of the invention unlessotherwise claimed. No language in the specification should be construedas indicating any non-claimed element as essential to the practice ofthe invention.

Preferred embodiments of this invention are described herein, includingthe best mode known to the inventors for carrying out the invention.Variations of those preferred embodiments may become apparent to thoseof ordinary skill in the art upon reading the foregoing description. Theinventors expect skilled artisans to employ such variations asappropriate, and the inventors intend for the invention to be practicedotherwise than as specifically described herein. Accordingly, thisinvention includes all modifications and equivalents of the subjectmatter recited in the claims appended hereto as permitted by applicablelaw. Moreover, any combination of the above-described elements in allpossible variations thereof is encompassed by the invention unlessotherwise indicated herein or otherwise clearly contradicted by context.

The invention claimed is:
 1. An isolated or purified nucleic acidsequence with at least 76.05% identity to SEQ ID NO:
 1. 2. The nucleicacid sequence of claim 1, which has at least 80% identity to SEQ IDNO:
 1. 3. The nucleic acid sequence of claim 1, which has at least 85%identity to SEQ ID NO:
 1. 4. The nucleic acid sequence of claim 1, whichhas at least 90% identity to SEQ ID NO:
 1. 5. The nucleic acid sequenceof claim 1, which has at least 95% identity to SEQ ID NO:
 1. 6. Thenucleic acid sequence of claim 1, which comprises SEQ ID NO:
 1. 7. Anisolated or purified nucleic acid sequence encoding an amino acidsequence with at least 97.15% identity to SEQ ID NO:
 2. 8. A nucleicacid sequence of claim 7, which encodes SEQ ID NO:
 2. 9. The nucleicacid sequence of claim 1, further comprising a nucleic acid sequencethat encodes ubiquitin.
 10. The nucleic acid sequence of claim 9, whichencodes SEQ ID NO:
 4. 11. The nucleic acid sequence of claim 10, whichcomprises SEQ ID NO:
 3. 12. An isolated or purified nucleic acidsequence with at least 82.56% identity to SEQ ID NO:
 5. 13. The nucleicacid sequence of claim 12, which has at least 90% identity to SEQ ID NO:5.
 14. The nucleic acid sequence of claim 12, which has at least 95%identity to SEQ ID NO:
 5. 15. The nucleic acid sequence of claim 12,which comprises SEQ ID NO:
 5. 16. A vector comprising the nucleic acidsequence of claim
 1. 17. A vector comprising (i) a nucleic acid sequencewith at least 74.5% identity to SEQ ID NO: 1 and (ii) a nucleic acidsequence with at least 82.56% identity to SEQ ID NO:
 5. 18. The vectorof claim 17, wherein the nucleic acid sequence (i) has at least 80%identity to SEQ ID NO:
 1. 19. The vector of claim 17, wherein thenucleic acid sequence (i) has at least 85% identity to SEQ ID NO:
 1. 20.The vector of claim 17, wherein the nucleic acid sequence (i) has atleast 90% identity to SEQ ID NO:
 1. 21. The vector of claim 17, whereinthe nucleic acid sequence (i) has at least 95% identity to SEQ ID NO: 1.22. The vector of claim 17, wherein the nucleic acid sequence (ii) hasat least 85% identity to SEQ ID NO:
 5. 23. The vector of claim 17,wherein the nucleic acid sequence (ii) has at least 90% identity to SEQID NO:
 5. 24. The vector of claim 17, wherein the nucleic acid sequence(ii) has at least 95% identity to SEQ ID NO:
 5. 25. The vector of claim17, wherein the vector comprises (i) SEQ ID NO: 1 and (ii) SEQ ID NO: 5.26. The vector of claim 17, wherein the vector is a viral vector or aplasmid.
 27. The vector of claim 26, wherein the vector is a poxvirus.28. The vector of claim 26, wherein the vector is a vaccinia virus. 29.The vector of claim 26, wherein the vector is an adeno-associatedvector.
 30. The vector of claim 26, wherein the vector is an adenoviralvector.
 31. The vector of claim 26, wherein the adenoviral vectorcomprises an adenoviral genome, and wherein a gorilla adenovirus is thesource of the adenoviral genome.
 32. The vector of claim 31, wherein thegorilla adenovirus comprises 70% or more identity to a nucleic acidsequence selected from the group consisting of SEQ ID NO: 9, SEQ ID NO:10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ IDNO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24,SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, andcombinations thereof.
 33. The vector of claim 32, wherein the gorillaadenovirus comprises 80% or more identity to a nucleic acid sequenceselected from the group consisting of SEQ ID NO: 9, SEQ ID NO: 10, SEQID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15,SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO:20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ IDNO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, and combinationsthereof.
 34. The vector of claim 33, wherein the gorilla adenoviruscomprises 90% or more identity to a nucleic acid sequence selected fromthe group consisting of SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16,SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO:21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ IDNO: 26, SEQ ID NO: 27, SEQ ID NO: 28, and combinations thereof.
 35. Thevector of claim 34, wherein the gorilla adenovirus comprises 95% or moreidentity to a nucleic acid sequence selected from the group consistingof SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO:13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ IDNO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27,SEQ ID NO: 28, and combinations thereof.
 36. The vector of claim 35,wherein the gorilla adenovirus comprises a nucleic acid sequenceselected from the group consisting of SEQ ID NO: 9, SEQ ID NO: 10, SEQID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15,SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO:20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ IDNO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, and combinationsthereof.
 37. The vector of claim 30, wherein the adenoviral vectorrequires complementation of a deficiency in one or more early regions ofthe adenoviral genome for propagation.
 38. The vector of claim 37,wherein the adenoviral vector requires complementation of a deficiencyin the E1 region of the adenoviral genome for propagation.
 39. Thevector of claim 37, wherein the adenoviral vector requirescomplementation of a deficiency in the E1A region of the adenoviralgenome for propagation.
 40. The vector of claim 37, wherein theadenoviral vector requires complementation of a deficiency in the E1Bregion of the adenoviral genome for propagation.
 41. The vector of claim37, wherein the adenoviral vector requires complementation of adeficiency in the E2 region of the adenoviral genome for propagation.42. The vector of claim 37, wherein the adenoviral vector requirescomplementation of a deficiency in the E4 region of the adenoviralgenome for propagation.
 43. The vector of claim 37, wherein theadenoviral vector requires complementation of a deficiency in the E1region of the adenoviral genome and a deficiency in the E4 region of theadenoviral genome for propagation.
 44. The vector of claim 37, whereinthe adenoviral vector requires complementation of a deficiency in the E1region of the adenoviral genome and a deficiency in the E2 region of theadenoviral genome for propagation.
 45. The vector of claim 37, whereinthe adenoviral vector further comprises a deletion of all or part of theE3 region of the adenoviral genome.
 46. A polypeptide encoded by thenucleic acid sequence of claim
 1. 47. A polypeptide with at least 98%identity to SEQ ID NO:
 2. 48. A composition comprising the nucleic acidsequence of claim 1 and a pharmaceutically acceptable carrier.
 49. Acomposition comprising the vector of claim 16 and a pharmaceuticallyacceptable carrier.
 50. A composition comprising the polypeptide ofclaim 46 and a pharmaceutically acceptable carrier.
 51. A method ofinducing an immune response against Herpes Simplex Virus (HSV) in amammal, which method comprises administering to the mammal thecomposition of claim 48, whereupon an immune response against HSV isinduced in the mammal.
 52. A method of treating or preventing a HerpesSimplex Virus (HSV) disease in a mammal, which method comprisesadministering to the mammal an effective amount of the composition ofclaim 49, whereupon a HSV disease is treated or prevented in the mammal.53. A method of inducing a T cell response against Herpes Simplex Virus(HSV) in a mammal, which method comprises administering to the mammalthe composition of claim 49, whereupon a T cell response against HSV isinduced in the mammal.
 54. The method of claim 53, wherein the T cellresponse is a mucosal T cell response.
 55. A method of inducing animmune response against Herpes Simplex Virus (HSV) in a mammal, whichmethod comprises administering to the mammal a first administration of acomposition and at least one additional administration of the same ordifferent composition, wherein each of the administrations comprises avector comprising: (a) a nucleic acid sequence with at least 76.05%identity to SEQ ID NO: 1, (b) a nucleic acid sequence encoding an aminoacid sequence with at least 97.15% identity to SEQ ID NO: 2, or (c) anucleic acid sequence with at least 82.56% identity to SEQ ID NO: 5, andwherein the vector in the compositions can be the same or different,whereupon an immune response against HSV is induced in the mammal. 56.The method of claim 55, wherein the immune response is a mucosal immuneresponse.
 57. A method of treating or preventing a Herpes Simplex Virus(HSV) disease in a mammal, which method comprises administering to themammal an effective amount of each of a first administration of acomposition and at least one additional administration of the same ordifferent composition, wherein each of the administrations comprises avector comprising: (a) a nucleic acid sequence with at least 76.05%identity to SEQ ID NO: 1, (b) a nucleic acid sequence encoding an aminoacid sequence with at least 97.15% identity to SEQ ID NO: 2, or (c) anucleic acid sequence with at least 82.56% identity to SEQ ID NO: 5, andwherein the vector in the administrations of compositions can be thesame or different, whereupon a HSV disease is treated or prevented inthe mammal.
 58. A method of inducing a T cell response against HerpesSimplex Virus (HSV) in a mammal, which method comprises administering tothe mammal a first administration of a composition and at least oneadditional administration of the same or different composition, whereineach of the administrations comprises a vector comprising: (a) a nucleicacid sequence with at least 76.05% identity to SEQ ID NO: 1, (b) anucleic acid sequence encoding an amino acid sequence with at least97.15% identity to SEQ ID NO: 2, or (c) a nucleic acid sequence with atleast 82.56% identity to SEQ ID NO: 5, and wherein the vector in theadministrations of compositions can be the same or different, whereupona T cell response against HSV is induced in the mammal.
 59. The methodof claim 58, wherein the T cell response is a mucosal T cell response.60. A method of inducing an antibody response against Herpes SimplexVirus (HSV) in a mammal, which method comprises administering to themammal a first administration of a composition and at least oneadditional administration of the same or different composition, whereineach of the administrations comprises the vector of claim 16, andwherein the vector in the administrations of compositions can be thesame or different, whereupon an antibody response against HSV is inducedin the mammal.
 61. A method of treating or preventing a Herpes SimplexVirus (HSV) disease in a mammal, which method comprises administering tothe mammal an effective amount of (a) and (b): (a) a first compositioncomprising the vector of claim 31, and (b) a second compositioncomprising the vector of claim 31, wherein (i) the second composition isadministered after the first composition, and (ii) the vector in thecompositions are the same or different, whereupon the T cell responseinduced after administration of the first and second compositions isgreater than the T cell response induced after administration of thefirst composition alone or the second composition alone, and a HSVdisease is treated or prevented in the mammal.
 62. The method of claim51, which further comprises administering one or more cytokines oradjuvants to the mammal.
 63. The method of claim 51, wherein the HSV isHSV-1.
 64. The method of claim 51, wherein the HSV is HSV-2.
 65. Themethod of claim 51, wherein the administration is intramuscularadministration, intradermal administration, or subcutaneousadministration.
 66. A vector comprising the nucleic acid sequence ofclaim
 8. 67. A polypeptide encoded by the nucleic acid sequence of claim8.
 68. A composition comprising the nucleic acid sequence of claim 8 anda pharmaceutically acceptable carrier.
 69. A composition comprising thevector of claim 66 and a pharmaceutically acceptable carrier.
 70. Acomposition comprising the polypeptide of claim 47 and apharmaceutically acceptable carrier.